CA2349818A1 - Membrane transport proteins - Google Patents

Abstract

The invention provides human membrane transport proteins (MTRP) and polynucleotides which identify and encode MTRP. The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists. The invention also provides methods for diagnosing, treating, or preventing disorders associated with expression of MTRP.

Description

MEMBRANE TRANSPORT PROTEINS
TECHNICAL FIELD
This invention relates to nucleic acid and amino acid sequences of membrane transport proteins and to the use of these sequences in the diagnosis, treatment, and prevention of membrane transport disorders; immune/inflammatory disorders; and cell proliferative disorders including cancer.
BACKGROUND OF THE INVENTION
Eukaryotic cells are bound by a lipid bilayer membrane and subdivided into functionally distinct, membrane bound compartments. The membranes maintain essential differences between the cytosol, the extracellular environment, and the contents of intracellular organelles such as the Golgi or the endoplasmic reticulum. As lipid membranes are highly impermeable to most polar molecules, transport of essential nutrients; metal ions such as K', NH,', P;, SOa'--;
sugars; vitamins; metabolic waste products; cell signaling molecules; drugs; peptides: and proteins and other macromolecules across lipid membranes and between organelles must be mediated by a variety of transport molecules.
Many transport mechanisms are substrate specific, with each transport protein carrying particular members of a molecular class, such as ions, sugars, or amino acids, across membranes. For example, amino acids are imported into cells via specific amino acid permeases.
Transport proteins are multi-pass transmembrane proteins, which either actively transport molecules across the membrane or passively allow them to cross. Active transport involves directional pumping of a solute across the membrane, usually against an electrochemical gradient.
Active transport is tightly coupled to a source of metabolic energy, such as ATP hydrolysis or an electrochemically favorable ion gradient. Passive transport involves the movement of a solute down its electrochemical gradient. Transport proteins can be further classified as either carrier proteins or channel proteins. Carrier proteins, which can function in active or passive transport, bind to a specific solute to be transported and undergo a conformational change which transfers the bound solute across the membrane. Channel proteins, which only function in passive transport, form hydrophilic pores across the membrane. When the pores open, specific solutes, such as inorganic ions, pass through the membrane and down the electrochemical gradient of the solute.
Transport proteins play roles in antibiotic resistance, toxin secretion, ion balance, synaptic neurotransmission, kidney function, intestinal absorption, tumor growth, and other diverse cell functions (Griffith, J. and C. Sansom (1998) The Transporter Facts Book, Academic Press, San Diego CA, pp. 3-29). A variety of human inherited diseases are caused by mutation of transport proteins.
For example, cystinuria is an inherited disease that results from the inability to transport cystine, the disulfide-linked dimer of cysteine, from the urine into the blood.
Accumulation of cystine in the urine leads to the formation of cystine stones in the kidneys. Also, many transport proteins are composed of subunits that may confer specificity for the tissue in which the transport mechanism functions, and are therefore associated with tissue-specific disorders. Examples of transport proteins include facilitative transporters, the secondary active symporters and antiporters driven by ion gradients, and active ATP binding cassette transporters involved in multiple-drug resistance and targeting of antigenic peptides to MHC Class 1 molecules, and the El-E2 cation transport ATPases.
Carrier proteins which transport a single solute from one side of the membrane to the other are called uniporters. In contrast, coupled transporters link the transfer of one solute with l0 simultaneous or sequential transfer of a second solute, either in the same direction (symport) or in the opposite direction (antiport). For example, intestinal and kidney epithelium contains a variety of symporter systems wherein the movement of sodium into the cell down its electrochemical gradient co-transports a second solute into the cell. The sodium gradient that provides the driving force for solute uptake is maintained by the ubiquitous Na;/K' ATPase. Sodium-coupled transporters include the mammalian glucose transporter (SGLT1 ), iodide transporter (NIS), and multivitamin transporter (SMVT). These three transporters have twelve putative transmembrane segments, extracellular glycosylation sites, and cytoplasmically-oriented N- and C-termini. NIS plays a crucial role in the evaluation, diagnosis, and treatment of various thyroid pathologies because it is the molecular basis for radioiodide thyroid-imaging techniques and for specific targeting of radioisotopes to the thyroid 2o gland (Levy, O. et al. ( 1997) Proc. Natl. Acad. Sci. USA 94:5568-5573).
SMVT is expressed in the intestinal mucosa, kidney, and placenta, and is implicated in the transport of the water-soluble vitamins, e.g., biotin and pantothenate (Prasad, P.D. et al. (1998) J. Biol.
Chem. 273:7501-7506).
The largest and most diverse family of transport proteins is the ATP-binding cassette (ABC) transporters. As a family, ABC transporters can transport substances that differ markedly in chemicat structure and size, ranging from small molecules such as ions, sugars, amino acids, peptides, and phospholipids, to lipopeptides, large proteins, and complex hydrophobic drugs.
Each ABC
transporter consists of four modules: two nucleotide-binding domains (NBDs),,which hydrolyze ATP
to supply the energy required for transport; and two membrane-spanning domains (MSDs), which may form membrane channels. The NBDs consist of approximately two hundred conserved amino acid residues while the MSDs each contain six putative transmembrane segments.
(See, e.g., Saurin, W. et al. (1994) Mol. Microbiol. 12:993-1004; Shani, N. et al. (1996) J. Biol.
Chem. 271:8725-8730;
Koster, W. and B. Bohm ( 1992) Mol. Gen. Genet. 232:399-407.) The four ABC
transporter modules may be encoded by a single gene, as is the case for the cystic fibrosis transmembrane conductance regulator (CFTR), or by separate genes. When encoded by separate genes, each gene product contains a single NBD and MSD. These "half molecules" form homo- and heterodimers, such as Tapl and Tap2, the endoplasmic reticulum-based major histocompatibility (MHC') peptide transport system associated with antigen processing (Androlewicz, M.J. et al. ( 1994) Proc. Natl. Acad. Sci.
USA 91:12716-12720).
Several genetic diseases are attributed to defects in ABC transporters, including the following diseases and their corresponding proteins: cystic fibrosis (CFTR, an ion channel; Welsh, M.J. and A.E. Smith (1993) Cell 73:1251-1254); X-linked adrenoleukodystrophy, an inborn error of peroxisomal ~i-oxidation of very long chain fatty acids (adrenoleukodystrophy protein, ALDP);
Zellweger syndrome, an inborn error of peroxisome biogenesis (peroxisomal membrane protein-70, PMP70); and hyperinsulinemic hypoglycemia (sulfonylurea receptor, SUR). The ABC transporters known as P-glycoproteins, or multidrug resistance (MDR) proteins; are associated with resistance to a wide range of hydrophobic drugs (MDR1; Gottesman, M.M. and I. Pastan (1993) Annu. Rev.
Biochem. 62:385-427) or with phosphatidylcholine transport (MDR2; Ruetz, S.
and P. Gros (1994) Cell 77:1071-1081 ). MDR is common in cancer cells, and contributes to low efficacy or failure of chemotherapy (Taglight, D. and S. Michaelis (1998) Methods Enzymol. 292:131-163). MDR is mediated by transporters, e.g., P-glycoproteins or the multidrug resistance-associated protein MRP, that normally function in the liver, intestines, and kidney to move toxic substances from the cytosol into the bile, intestinal lumen, or urine. In cancerous cells, these transporters extrude chemotherapeutic agents into the extracellular space, thereby conferring drug resistance. Recently, an ABC transporter-type protein was isolated from a human leukemia cell line.
This transporter, termed the anthracycline resistance associated protein (GI 1279457, SEQ ID N0:42), is overexpressed in a multidrug resistant leukemia cell sub-line, and has sequence homology with other multidrug-resistance associated proteins including MRP (Longhurst, T.J. et al. (1996) Br. J. Cancer 74:1331-1335).
Transport of fatty acids across the plasma membrane can occur by diffusion, a high capacity, low affinity process. However, under normal physiological conditions a significant fraction of fatty acid transport appears to occur via a high affinity, low capacity protein-mediated transport process.
Fatty acid transport protein (FATP), an integral membrane protein with four transmembrane segments, is expressed in tissues exhibiting high levels of plasma membrane fatty acid flux, such as muscle, heart, and adipose. Expression of FATP is upregulated in 3T3-L1 cells during adipose conversion, and expression in COS7 fibroblasts elevates the cells' uptake of long-chain fatty acids.
Expression studies suggest a role for FATP in lipid metabolism, obesity, and type II diabetes mellitus (Hui, T.Y. et al. (1998) J. Biol. Chem. 273:27420-27429).
EI-E2 (or P-type) ATPases constitute a superfamily of cation transporters present in both prokaryotes and eukaryotes that mediate membrane flux of all biologically relevant cations. These ATPases are postulated to exist in two different conformational states, designated EI and E2, during the course of the ATP hydrolysis reaction, and to conserve the energy from ATP
hydrolysis in the form of an acyl phosphate, primarily an aspartyl phosphate. Members of this family are divided into four major groups; the Ca+Z-transporting ATPases, Na+/K+ -and gastric H+/K+-transporting ATPases, plasma membrane H+-transporting ATPases (proton pumps), and the bacterial P-type ATPases (BLOCKS: BL00154, P-type cation-transporting ATPase superfamily signature).
The metabolism of amino acids is complex and highly regulated. While cells are capable of creating most amino acids de novo, the import of amino acids into cells via specific amino acid permease proteins is vital for maintaining the appropriate and complete availability of all necessary amino acids. This is particularly important during cell proliferation and differentiation. In addition to their role as protein building blocks, amino acids also serve as precursors for a variety of other important macromolecules. For example, the hormone thyroxine, the pigment melanin, and the neurotransmitters histamine, epinephrine, and serotonin are produced from various amino acid IS precursors, including histidine, tyrosine, and tryptophan. A component of sphingolipid formation, sphingosine, is derived from serine. Porphyrin rings, which are components of heme molecules, use glycine as a nitrogen donor. Significant portions of the ring structures of purines and pyrimidines, components of nucleic acids, are formed from the breakdown of numerous amino acids. Amino acids are also important in energy metabolism. Unlike fatty acids and glucose, amino acids cannot be stored in the cell, so excess amino acids are fed into the citric acid cycle to produce energy molecules including fatty acids, ketone bodies, and glucose. Thus, precise control of amino acid metabolism is extremely important to both proliferating and non-proliferating cells.
The E16 gene, cloned from human peripheral blood lymphocytes, encodes a 241 amino acid integral membrane protein with multiple predicted transmembrane domains (Gaugitsch, H.W. et al.
(1992) J. Biol. Chem. 267:11267-11273). E16 gene expression is closely linked to cellular activation and division. In myeloid and lymphoid cells, E16 transcripts are rapidly induced and rapidly degraded after stimulation. This pattern of expression resembles the kinetics seen for proto-oncogenes and lymphokines in the T cell system. Elevated levels of E16 expression were detected in colonic, gastric, and breast adenocarcinomas, and in lymphoma, while little or no E16 expression was detected in normal (non-cancerous) human tissues such as adult brain, lung, liver, colon, esophagus, stomach, or kidney, nor in four-month fetal brain, lung, liver, or kidney (Wolf, D.A. et al. (1996) Cancer Res. 56:5012-5022; Gaugitsch et al., supra). E16 was detected in every cell line tested. Its presence in rapidly dividing cell lines and its absence in human tissues with low proliferative potential suggest that E16 is directly involved in the cell division process, where it helps provide important building blocks for energy metabolism, biochemical synthetic pathways, and protein synthesis.
Post-translationa) modification of polypeptides occurs in the lumen of the' Golgi apparatus.
Such modifications include, for example, the addition of sugar molecules by enzymes such as N-acetylglucosaminyltransferase, to produce glycoproteins. The sugar-donating molecules in this reaction are typically nucleotide sugars, such as uridine diphosphate-galactose (UDP-Gal). UPD-Gal and other nucleotide sugars are transported from the cytosol into the Golgi apparatus by specific transporter molecules. The availability of these nucleotide sugars can regulate which glycoproteins are synthesized, and therefore has a significant impact on cellular function (Toms, L. et al. (1996) J.
Biol. Chem. 271:3897-3901; Guillen, E. et al. (1998) Proc. Natl. Acad. Sci.
USA 95:7888-7892).
The discovery of new membrane transport proteins and the polynucleotides encoding them satisfies a need in the art by providing new compositions which are useful in the diagnosis, prevention, and treatment of membrane transport disorders; immune/inflammatory disorders; and cell proliferative disorders including cancer.
l5 SUMMARY OF THE INVENTION
The invention features substantially purified polypeptides, membrane transport proteins, referred to collectively as "MTRP" and individually as "MTRP-I," "MTRP-2,"
"MTRP-3," "MTRP-4," "MTRP-S," "MTRP-6," "MTRP-7," "MTRP-8," "MTRP-9," "MTRP-10," "MTRP-11,"
"MTRP-12," "MTRP-13," "MTRP-14," "MTRP-15," "MTRP-16," and "MTRP-17." In one aspect, the invention provides a substantially purified polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-17 and fragments thereof. The invention also includes a polypeptide comprising an amino acid sequence that differs by one or more conservative amino acid substitutions from an amino acid sequence selected from the group consisting of SEQ ID NO:1-17.
The invention further provides a substantially purified variant having at least 90% amino acid identity to at least one of the amino acid sequences selected from the group consisting of SEQ ID
NO:1-17 and fragments thereof. The invention also provides an isolated and purified polynucleotide encoding the polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-17 and fragments thereof. The invention also includes an isolated and purified polynucleotide variant having at least 90% polynucleotide sequence identity to the poiynucleotide encoding the polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:I-17 and fragments thereof.
Additionally, the invention provides an isolated and purified polynucleotide which hybridizes under stringent conditions to the polynucleotide encoding the polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-17 and fragments thereof. The invention also provides-an isolated and purified polynucleotide having a sequence which is complementary to the polynucleotide encoding the polypeptide comprising the amino acid sequence selected from the group consisting of SEQ ID NO:1-17 and fragments thereof.
The invention also provides a method for detecting a polynucleotide in a sample containing nucleic acids, the method comprising the steps of: (a) hybridizing the complement of the polynucleotide sequence to at least one of the polynucleotides of the sample, thereby forming a hybridization complex; and (b) detecting the hybridization complex, wherein the presence of the hybridization complex correlates with the presence of a polynucleotide in the sample. In one aspect, the method further comprises amplifying the polynucleotide prior to hybridization.
I0 The invention also provides an isolated and purified polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO:I 8-34 and fragments thereof. The invention further provides an isolated and purified polynucleotide variant having at least 90% polynucleotide sequence identity to the polynucleotide sequence selected from the group consisting of SEQ ID N0:18-34 and fragments thereof. The invention also provides an isolated and I S purified polynucleotide having a sequence which is complementary to the polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ ID N0:18-34 and fragments thereof.
The invention further provides an expression vector containing at least a fragment of the polynucleotide encoding the polypeptide comprising an amino acid sequence selected from the group 20 consisting of SEQ ID NO:I-17. In another aspect, the expression vector is contained within a host cell.
The invention also provides a method for producing a polypeptide, the method comprising the steps of: (a) culturing the host cell containing an expression vector containing a polynucleotide of the invention under conditions suitable for the expression of the polypeptide; and (b) recovering the 25 polypeptide from the host cell culture.
The invention also provides a pharmaceutical composition comprising a substantially purified polypeptide having the amino acid sequence selected from the group consisting of SEQ ID NO:1-17 and fragments thereof, in conjunction with a suitable pharmaceutical carrier.
The invention further includes a purified antibody which binds to a polypeptide selected from 30 the group consisting of SEQ ID NO:1-17 and fragments thereof. The invention also provides a purified agonist and a purified antagonist to the polypeptide.
The invention also provides a method for treating or preventing a disorder associated with decreased expression or activity of MTRP, the method comprising administering to a subject in need of such treatment an effective amount of a pharmaceutical composition comprising a substantially purified polypeptide having the amino acid~sequence selected from the group consisting of SEQ ID
NO:1-17 and fragments thereof, in conjunction with a suitable pharmaceutical carrier.
The invention also provides a method for treating or preventing a disorder associated with increased expression or activity of MTRP, the method comprising administering to a subject in need of such treatment an effective amount of an antagonist of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-17 and fragments thereof.
BRIEF DESCRIPTION OF THE FIGURES AND TABLES
Figures lA, 1B, 1C, and 1D show the amino acid sequence alignment between MTRP-(Incyte Clone ID 1720440; SEQ ID N0:3) and mouse fatty acid transport protein (GI 2612939; SEQ
ID N0:35), produced using the multisequence alignment program of LASERGENE
software (DNASTAR, Madison Wl).
Figures 2A, 2B, 2C, and 2D show the amino acid sequence alignment between MTRP-(lncyte Clone ID 2274290; SEQ ID N0:4) and Schistosoma mansoni SMDR1 (GI
425474; SEQ ID
N0:36), produced using the multisequence alignment program of LASERGENE
software (DNASTAR).
Figures 3A, 3B, 3C, and 3D show the amino acid sequence alignment between MTRP-(Incyte Clone ID 2740029; SEQ ID NO:S) and rat sodium-dependent multivitamin transporter (GI
3015617; SEQ ID N0:37), produced using the multisequence alignment program of LASERGENE
software (DNASTAR).
Table 1 shows poiypeptide and nucleotide sequence identification numbers (SEQ
1D NOs), clone identification numbers (clone IDs), cDNA libraries, and cDNA fragments used to assemble full-length sequences encoding MTRP.
Table 2 shows features of each polypeptide sequence, including potential motifs, homologous sequences, and methods, algorithms, and searchable databases used for analysis of MTRP.
Table 3 shows the tissue-specific expression patterns of each nucleic acid sequence as determined by northern analysis; diseases, disorders, or conditions associated with these tissues; and the vector into which each cDNA was cloned.
Table 4 describes the tissues used to construct the cDNA libraries from which cDNA clones encoding MTRP were isolated.
Table 5 shows the tools, programs, and algorithms used to analyze MTRP, along with applicable descriptions, references, and threshold parameters.
DESCRIPTION OF THE INVENTION

Before the present proteins, nucleotide sequences, and methods are described, it is understood that this invention is not limited to the particular machines, materials and methods described, as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims.
It must be noted that as used herein and in the appended claims, the singular forms "a," "an,"
and "the" include plural reference unless the context clearly dictates otherwise. Thus, for example, a reference to "a host cell" includes a plurality of such host cells, and a reference to "an antibody" is a reference to one or more antibodies and equivalents thereof known to those skilled in the art, and so forth.
Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs.
Although any machines, materials, and methods similar or equivalent to those described herein can be used to practice or test the present invention, the preferred machines, materials and methods are now described. Al) publications mentioned herein are cited for the purpose of describing and disclosing the cell lines, protocols, reagents and vectors which are reported in the publications and which might be used in connection with the invention. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.
DEFINITIONS
"MTRP" refers to the amino acid sequences of substantially purified MTRP
obtained from any species, particularly a mammalian species, including bovine, ovine, porcine, murine, equine, and human, and from any source, whether natural, synthetic, semi-synthetic, or recombinant.
The term "agonist" refers to a molecule which intensifies or mimics the biological activity of MTRP: Agonists may include proteins, nucleic acids, carbohydrates, small molecules, or any other compound or composition which modulates the activity of MTRP either by directly interacting with MTRP or by acting on components of the biological pathway in which MTRP
participates.
An "allelic variant" is an alternative form of the gene encoding MTRP. Allelic variants may result from at least one mutation in the nucleic acid sequence and may result in altered mRNAs or in polypeptides whose structure or function may or may not be altered. A gene may have none, one, or many allelic variants of its naturally occurring form. Common mutational changes which give rise to allelic variants are generally ascribed to natural deletions, additions, or substitutions of nucleotides.
Each of these types of changes may occur alone, or in combination with the others, one or more times in a given sequence.
"Altered" nucleic acid sequences encoding MTRP include those sequences with deletions, insertions, or substitutions of different nucleotides, resulting in a polypeptide the same as MTRP or a polypeptide with at least one functional characteristic of MTRP. Included within this definition are polymorphisms which may or may not be readily detectable using a particular oligonucleotide probe of the polynucleotide encoding MTRP, and improper or unexpected hybridization to allelic variants, with a locus other than the normal chromosomal locus for the polynucleotide sequence encoding MTRP. The encoded protein may also be "altered," and may contain deletions, insertions, or substitutions of amino acid residues which produce a silent change and result in a functionally equivalent MTRP. Deliberate amino acid substitutions may be made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and/or the amphipathic nature of the residues, as long as the biological or immunological activity of MTRP is retained. For example, negatively charged amino acids may include aspartic acid and glutamic acid, and positively charged amino acids may include lysine and arginine. Amino acids with uncharged polar side chains having similar hydrophilicity values may include: asparagine and glutamine; and serine and threonine.
Amino acids with uncharged side chains having similar hydrophilicity values may include: leucine, 1 S isoleucine, and valine; glycine and alanine; and phenylalanine and tyrosine.
The terms "amino acid" and "amino acid sequence" refer to an oligopeptide, peptide, polypeptide, or protein sequence, or a fragment of any of these, and to naturally occurring or synthetic molecules. Where "amino acid sequence" is recited to refer to an amino acid sequence of a naturally occurring protein molecule, "amino acid sequence" and like terms are not meant to limit the amino acid sequence to the complete native amino acid sequence associated with the recited protein molecule.
"Amplification" relates to the production of additional copies of a nucleic acid sequence.
Amplification is generally carried out using polymerase chain reaction (PCR) technologies well known in the art.
The term "antagonist" refers to a molecule which inhibits or attenuates the biological activity of MTRP. Antagonists may include proteins such as antibodies, nucleic acids, carbohydrates, small molecules, or any other compound or composition which modulates the activity of MTRP either by directly interacting with MTRP or by acting on components of the biological pathway in which MTRP
participates.
The term "antibody" refers to intact immunoglobulin molecules as well as to fragments thereof, such as Fab, F(ab'),, and Fv fragments, which are capable of binding an epitopic determinant.
Antibodies that bind MTRP polypeptides can be prepared using intact polypeptides or using fragments containing small peptides of interest as the immunizing antigen. The polypeptide or oligopeptide used to immunize an animal (e.g., a mouse, a rat, or a rabbit) can be derived from the translation of RNA, or synthesized chemically, and can be conjugated to a carrier protein if desired.
Commonly used carriers that are chemically coupled to peptides include bovine serum albumin, thyroglobulin, and keyhole limpet hemocyanin (KLH). The coupled peptide is then used to immunize the animal.
The tenor "antigenic determinant" refers to that region of a molecule (i.e., an epitope) that makes contact with a particular antibody. When a protein or a fragment of a protein is used to immunize a host animal, numerous regions of the protein may induce the production of antibodies which bind specifically to antigenic determinants (particular regions or three-dimensional structures on the protein). An antigenic determinant may compete with the intact antigen (i.e., the immunogen used to elicit the immune response) for binding to an antibody.
The term "antisense" refers to any composition containing a nucleic acid sequence which is complementary to the "sense" strand of a specific nucleic acid sequence.
Antisense molecules may be produced by any method including synthesis or transcription. Once introduced into a cell, the complementary nucleotides combine with natural sequences produced by the cell to form duplexes and to block either transcription or translation. The designation "negative"
or "minus" can refer to the antisense strand, and the designation "positive" or "plus" can refer to the sense strand.
The term "biologically active" refers to a protein having structural, regulatory, or biochemical functions of a naturally occurring molecule. Likewise, "immunologically active" refers to the capability of the natural, recombinant, or synthetic MTRP, or of any oligopeptide thereof, to induce a specific immune response in appropriate animals or cells and to bind with specific antibodies.
The terms "complementary" and "complementarily" refer to the natural binding of polynucleotides by base pairing. For example, the sequence "5' A-G-T 3"' bonds to the complementary sequence "3' T-C-A S'." Complementarily between two single-stranded molecules may be "partial," such that only some of the nucleic acids bind, or it may be "complete," such that total complementarily exists between the single stranded molecules. The degree of complementarily between nucleic acid strands has significant effects on the efficiency and strength of the hybridization between the nucleic acid strands. This is of particular importance in amplification reactions, which depend upon binding between nucleic acid strands, and in the design and use of peptide nucleic acid (PNA) molecules.
A "composition comprising a given polynucleotide sequence" and a "composition comprising a given amino acid sequence" refer broadly to any composition containing the given polynucleotide or amino acid sequence. The composition may comprise a dry formulation or an aqueous solution.
Compositions comprising polynucleotide sequences encoding MTRP or fragments of MTRP may be employed as hybridization probes. The probes may be stored in freeze-dried form and may be associated with a stabilizing agent such as a carbohydrate.. In hybridizations, the probe may be deployed in an aqueous solution containing salts (e.g., NaCI), detergents (e.g., sodium dodecyl sulfate; SDS), and other components (e.g., Denhardt's solution, dry milk, salmon sperm DNA, etc.).
"Consensus sequence" refers to a nucleic acid sequence which has been resequenced to resolve uncalled bases, extended using the XL-PCR kit (Perkin-Elmer, Norwalk CT) in the S' and/or the 3' direction, and resequenced, or which has been assembled from the overlapping sequences of one or more Incyte Clones and, in some cases, one or more public domain ESTs, using a computer program for fragment assembly, such as the GELVIEW fragment assembly system (GCG, Madison WI). Some sequences have been both extended and assembled to produce the consensus sequence.
"Conservative amino acid substitutions" are those substitutions that, when made, least interfere with the properties of the original protein, i.e., the structure and especially the function of the protein is conserved and not significantly changed by such substitutions. The table below shows amino acids which may be substituted for an original amino acid in a protein and which are regarded as conservative amino acid substitutions.
Original Residue Conservative Substitution Ala Gly, Ser Arg His, Lys Asn Asp, Gln, His Asp Asn, Glu Cys Ala, Ser Gln Asn, Glu, His Glu Asp, Gin, His Gly Ala His Asn, Arg, Gln, Glu Ile Leu, Val Leu Ile, Val Lys Arg, Gln, Glu Met Leu, Ile Phe His, Met, Leu, Trp, Tyr Ser Cys, Thr Thr Ser, Val Trp Phe, Tyr Tyr His, Phe, Trp Val Ile, Leu, Thr Conservative amino acid substitutions generally maintain (a) the structure of the polypeptide backbone in the area of the substitution, for example, as a beta sheet or alpha helical conformation, (b) the charge or hydrophobicity of the molecule at the site of the substitution, and/or (c) the bulk of the side chain.
A "deletion" refers to a change in the amino acid or nucleotide sequence that results in the absence of one or more amino acid residues or nucleotides.

The term "derivative" refers to the chemical modification of a polypeptide sequence, or a polynucleotide sequence. Chemical modifications of a polynucleotide sequence c'an include, for example, replacement of hydrogen by an alkyl, acyl, hydroxyl, or amino group.
A derivative polynucleotide encodes a polypeptide which retains at least one biological or immunological function of the natural molecule. A derivative polypeptide is one modified by glycosylation, pegylation, or any similar process that retains at least one biological or immunological function of the polypeptide from which it was derived.
A "fragment" is a unique portion of MTRP or the polynucleotide encoding MTRP
which is identical in sequence to but shorter in length than the parent sequence. A
fragment may comprise up to the entire length of the defined sequence, minus one nucleotide/amino acid residue. For example, a fragment may comprise from S to 1000 contiguous nucleotides or amino acid residues. A fragment used as a probe, primer, antigen, therapeutic molecule, or for other purposes, may be at least 5, 10, 15, 20, 25, 30, 40, 50, 60, 75, 100, 150, 250 or at least 500 contiguous nucleotides or amino acid residues in length. Fragments may be preferentially selected from certain regions of a molecule. For example, a polypeptide fragment may comprise a certain length of contiguous amino acids selected from the first 250 or 500 amino acids (or first 25% or SO% of a polypeptide) as shown in a certain defined sequence. Clearly these lengths are exemplary, and any length that is supported by the specification, including the Sequence Listing, tables, and figures, may be encompassed by the present embodiments.
A fragment of SEQ ID NO:I 8-34 comprises a region of unique polynucleotide sequence that specifically identifies SEQ ID N0:18-34, for example, as distinct from any other sequence in the same genome. A fragment of SEQ ID N0:18-34 is useful, for example, in hybridization and amplification technologies and in analogous methods that distinguish SEQ ID
NO:I 8-34 from related polynucleotide sequences. The precise length of a fragment of SEQ ID N0:18-34 and the region of SEQ ID N0:18-34 to which the fragment corresponds are routinely determinable by one of ordinary skill in the art based on the intended purpose for the fragment.
A fragment of SEQ ID NO:I-l7 is encoded by a fragment of SEQ ID N0:18-34. A
fragment of SEQ ID NO:I-17 comprises a region of unique amino acid sequence that specifically identifies SEQ 1D NO:I-17. For example, a fragment of SEQ ID NO:1-17 is useful as an immunogenic peptide for the development of antibodies that specifically recognize SEQ ID NO:1-17.
The precise length of a fragment of SEQ ID NO:1-17 and the region of SEQ ID NO:1-17 to which the fragment corresponds are routinely determinable by one of ordinary skill in the art based on the intended purpose for the fragment.
The term "similarity" refers to a degree of complementarity. There may be partial similarity or complete similarity. The word "identity" may substitute for the word "similarity." A partially complementary sequence that at least partially inhibits an identical sequence from hybridizing to a target nucleic acid is referred to as "substantially similar." The inhibition of hybridization of the completely complementary sequence to the target sequence may be examined using a hybridization assay (Southern or northern blot, solution hybridization, and the like) under conditions of reduced stringency. A substantially similar sequence or hybridization probe will compete for and inhibit the binding of a completely similar (identical) sequence to the target sequence under conditions of reduced stringency. This is not to say that conditions of reduced stringency are such that non-specific binding is permitted, as reduced stringency conditions require that the binding of two sequences to one another be a specific (i.e., a selective) interaction. The absence of non-specific binding may be tested by the use of a second target sequence which lacks even a partial degree of complementarity (e.g., less than about 30% similarity or identity). In the absence of non-specific binding, the substantially similar sequence or probe will not hybridize to the second non-complementary target sequence.
The phrases "percent identity" and "% identity," as applied to polynucleotide sequences, refer to the percentage of residue matches between at least two polynucleotide sequences aligned using a standardized algorithm. Such an algorithm may insert, in a standardized and reproducible way, gaps in the sequences being compared in order to optimize alignment between two sequences, and therefore achieve a more meaningful comparison of the two sequences.
Percent identity between polynucleotide sequences may be determined using the default parameters of the CLUSTAL V algorithm as incorporated into the MEGALIGN
version 3.12e sequence alignment program. This program is part of the LASERGENE software package, a suite of molecular biological analysis programs (DNASTAR, Madison WI). CLUSTAL V is described in Higgins, D.G. and P.M. Sharp (1989) CABIOS 5:151-153 and in Higgins, D.G. et al. (1992) CABIOS
8:189-191. For pairwise alignments of polynucleotide sequences, the default parameters are set as follows: Ktuple=2, gap penalty=5, window=4, and "diagonals saved"=4. The "weighted" residue weight table is selected as the default. Percent identity is reported by CLUSTAL V as the "percent similarity" between aligned polynucleotide sequence pairs.
Alternatively, a suite of commonly used and freely available sequence comparison algorithms is provided by the National Center for Biotechnology Information (NCBI) Basic Local Alignment Search Tool (BLAST) (Altschul, S.F. et al. (1990) J. Mol. Biol. 215:403-410), which is available from several sources, including the NCBI, Bethesda, MD, and on the Internet at http://ww<v.ncbi.nlm.nih.gov/BLAST/. The BLAST software suite includes various sequence analysis programs including "blastn," that is used to align a known polynucleotide sequence with other polynucleotide sequences from a variety of databases. Also available is a tool called "BLAST 2 Sequences" that is used-for direct pairwise comparison of two nucleotide sequences. "BLAST 2 Sequences" can be accessed and used interactively at http://www.ncbi.nlm.nih.go'v/gorf/bl2.html. The "BLAST 2 Sequences" tool can be used for both blastn and blastp (discussed below). BLAST
programs are commonly used with gap and other parameters set to default settings. For example, to compare two nucleotide sequences, one may use blastn with the "BLAST 2 Sequences" tool Version 2Ø9 (May-07-1999) set at default parameters. Such default parameters may be, for example:
Matrix: BLOSUM62 Reward for match: I
Penalty for mismatch: -2 Open Gap: S and Extension Gap: 2 penalties Gap x drop-off SO
Expect: 1 D
Word Size: I1 Filter: on Percent identity may be measured over the length of an entire defined sequence, for example, as defined by a particular SEQ ID number, or may be measured over a shorter length, for example, over the length of a fragment taken from a larger, defined sequence, for instance, a fragment of at least 20, at least 30, at least 40, at least 50, at least 70, at least 100, or at least 200 contiguous nucleotides. Such lengths are exemplary only, and it is understood that any fragment length supported by the sequences shown herein, in the tables, figures, or Sequence Listing, may be used to describe a length over which percentage identity may be measured.
Nucleic acid sequences that do not show a high degree of identity may nevertheless encode similar amino acid sequences due to the degeneracy of the genetic code. It is understood that changes in a nucleic acid sequence can be made using this degeneracy to produce multiple nucleic acid sequences that all encode substantially the same protein.
The phrases "percent identity" and "% identity," as applied to polypeptide sequences, refer to the percentage of residue matches between at least two polypeptide sequences aligned using a standardized algorithm. Methods of polypeptide sequence alignment are well-known. Some alignment methods take into account conservative amino acid substitutions.
Such conservative substitutions, explained in more detail above, generally preserve the hydrophobicity and acidity at the site of substitution, thus preserving the structure (and therefore function) of the polypeptide.
Percent identity between polypeptide sequences may be determined using the default parameters of the CLUSTAL V algorithm as incorporated into the MEGALIGN
version 3.12e sequence alignment program (described and referenced above). For pairwise alignments of polypeptide sequences using CLUSTAL V, the default parameters are set as follows: Ktuple=1, gap penalty=3, window=5, and "diagonals saved"=5. The PAM250 matrix is selected as the default residue weight table. As with polynucleotide alignments, the percent identity is reported by CLUSTAL V as the "percent similarity" between aligned polypeptide sequence pairs.
Alternatively the NCBI BLAST software suite may be used. For example, for a pairwise comparison of two polypeptide sequences, one may use the "BLAST 2 Sequences"
tool Version 2Ø9 (May-07-1999) with blastp set at default parameters. Such default parameters may be, for example:
Matrix: BLOSUM62 Open Gap: 11 and Extension Gap: 1 penalties Gap x drop-off. 50 Expect. 10 Word Size: 3 Filter.' on Percent identity may be measured over the length of an entire defined polypeptide sequence, for example, as defined by a particular SEQ ID number, or may be measured over a shorter length, for example, over the length of a fragment taken from a larger, defined polypeptide sequence, for instance, a fragment of at least 15, at least 20, at least 30, at Least 40, at least 50, at least 70 or at least 150 contiguous residues. Such lengths are exemplary only, and it is understood that any fragment length supported by the sequences shown herein, in the tables, figures or Sequence Listing, may be used to describe a length over which percentage identity may be measured.
"Human artificial chromosomes" (HACs) are linear microchromosomes which may contain DNA sequences of about 6 kb to 10 Mb in size, and which contain all of the elements required for stable mitotic chromosome segregation and maintenance.
The term "humanized antibody" refers to antibody molecules in which the amino acid sequence in the non-antigen binding regions has been altered so that the antibody more closely resembles a human antibody, and still retains its original binding ability.
"Hybridization" refers to the process by which a polynucleotide strand anneals with a complementary strand through base pairing under defined hybridization conditions. Specific hybridization is an indication that two nucleic acid sequences share a high degree of identity. Specific hybridization complexes form under permissive annealing conditions and remain hybridized after the "washing" step(s). The washing steps) is particularly important in determining the stringency of the hybridization process, with more stringent conditions allowing less non-specific binding, i.e., binding between pairs of nucleic acid strands that are not perfectly matched.
Permissive conditions for annealing of nucleic acid sequences are routinely determinable by one of ordinary skill in the art and may be consistent among hybridization experiments, whereas wash conditions may be varied among experiments to achieve the desired stringency, and therefore hybridization specificity. Permissive annealing conditions occur, for example, at 68°C in the presence of about 6 x SSC, about 1 % (w/v) SDS, and about 100 pg/ml denatured salmon sperm DNA.
Generally, stringency of hybridization is expressed, in part, with reference to the temperature under which the wash step is carried out. Generally, such wash temperatures are selected to be about S°C to 20°C lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength and pH. The Tm is the temperature (under defined ionic strength and pH) at which SO% of the target sequence hybridizes to a perfectly matched probe. An equation for calculating T," and conditions for nucleic acid hybridization are well known and can be found in Sambrook et al., 1989, Molecular Cloning: A Laboratory Manual, 2"° ed., vol. 1-3, Cold Spring Harbor Press, Plainview NY;
specifically see volume 2, chapter 9.
High stringency conditions for hybridization between polynucleotides of the present invention include wash conditions of 68°C in the presence of about 0.2 x SSC and about 0.1 % SDS, for 1 hour.
Alternatively, temperatures of about 65°C, 60°C, 55°C, or 42°C may be used. SSC concentration may be varied from about 0.1 to 2 x SSC, with SDS being present at about 0.1 %. Typically, blocking reagents are used to block non-specific hybridization. Such blocking reagents include, for instance, denatured salmon sperm DNA at about 100-200 pg/ml. Organic solvent, such as formamide at a concentration of about 35-50% v/v, may also be used under particular circumstances, such as for RNA:DNA hybridizations. Useful variations on these wash conditions will be readily apparent to those of ordinary skill in the art. Hybridization, particularly under high stringency conditions, may be suggestive of evolutionary similarity between the nucleotides. Such similarity is strongly indicative of a similar role for the nucleotides and their encoded polypeptides.
The term "hybridization complex" refers to a complex formed between two nucleic acid sequences by virtue of the formation of hydrogen bonds between complementary bases. A
hybridization complex may be formed in solution (e.g., C°t or Rot analysis) or formed between one nucleic acid sequence present in solution and another nucleic acid sequence immobilized on a solid support (e.g., paper, membranes, filters, chips, pins or glass slides, or any other appropriate substrate to which cells or their nucleic acids have been fixed).
The words "insertion" and "addition" refer to changes in an amino acid or nucleotide sequence resulting in the addition of one or more amino acid residues or nucleotides, respectively.
"Immune response" can refer to conditions associated with inflammation, trauma, immune disorders, or infectious or genetic disease, etc. These conditions can be characterized by expression of various factors, e.g., cytokines, chemokines, and other signaling molecules, which may affect WO 00!26245 PCT/US99/26048 cellular and systemic defense systems.
The term "microarray" refers to an arrangement of distinct polynucleotide"s on a substrate.
The terms "element" and "array element" in a microarray context, refer to hybridizable polynucleotides arranged on the surface of a substrate.
The term "modulate" refers to a change in the activity of MTRP. For example, modulation may cause an increase or a decrease in protein activity, binding characteristics, or any other biological, functional, or immunological properties of MTRP.
The phrases "nucleic acid" and "nucleic acid sequence" refer to a nucleotide, oligonucleotide, polynucleotide, or any fragment thereof. These phrases also refer to DNA or RNA of genomic or synthetic origin which may be single-stranded or double-stranded and may represent the sense or the antisense strand, to peptide nucleic acid (PNA), or to any DNA-like or RNA-like material.
"Operably linked" refers to the situation in which a first nucleic acid sequence is placed in a functional relationship with the second nucleic acid sequence. For instance, a promoter is operably linked to a coding sequence if the promoter affects the transcription or expression of the coding IS sequence. Generally, operably linked DNA sequences may be in close proximity or contiguous and, where necessary to join two protein coding regions, in the same reading frame.
"Peptide nucleic acid" (PNA) refers to an antisense molecule or anti-gene agent which comprises an oligonucleotide of at least about 5 nucleotides in length linked to a peptide backbone of amino acid residues ending in lysine. The terminal lysine confers solubility to the composition.
PNAs preferentially bind complementary single stranded DNA or RNA and stop transcript elongation, and may be pegylated to extend their lifespan in the cell.
"Probe" refers to nucleic acid sequences encoding MTRP, their complements, or fragments thereof, which are used to detect identical, allelic or related nucleic acid sequences. Probes are isolated oligonucleotides or polynucleotides attached to a detectable label or reporter molecule.
Typical labels include radioactive isotopes, ligands, chemiluminescent agents, and enzymes.
"Primers" are short nucleic acids, usually DNA oligonucleotides, which may be annealed to a target polynucleotide by complementary base-pairing. The primer may then be extended along the target DNA strand by a DNA polymerase enzyme. Primer pairs can be used for amplification (and identification) of a nucleic acid sequence, e.g., by the polymerase chain reaction (PCR).
Probes and primers as used in the present invention typically comprise at least IS contiguous nucleotides of a known sequence. In order to enhance specificity, longer probes and primers may also be employed, such as probes and primers that comprise at least 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, or at least 150 consecutive nucleotides of the disclosed nucleic acid sequences. Probes and primers may be considerably longer than these examples, and it is understood that any length supported by the specification, includingthe tables, figures, and Sequence Listing, may be used.
Methods for preparing and using probes and primers are described in the references, for example Sambrook et al., 1989, Molecular Clonine: A Laboratory Manual, 2"d ed., vol. 1-3, Cold Spring Harbor Press, Plainview NY; Ausubel et al.,1987, Current Protocols in Molecular BioloQV, Greene Publ. Assoc. & Wiley-Intersciences, New York NY; Innis et al., 1990, PCR Protocols. A
Guide to Methods and Applications, Academic Press, San Diego CA. PCR primer pairs can be derived from a known sequence, for example, by using computer programs intended for that purpose such as Primer (Version 0.5, 1991, Whitehead Institute for Biomedical Research, Cambridge MA).
Oligonucleotides for use as primers are selected using software known in the art for such purpose. For example, OLIGO 4.06 software is useful for the selection of PCR
primer pairs of up to 100 nucleotides each, and for the analysis of oligonucleotides and larger polynucleotides of up to 5,000 nucleotides from an input polynucleotide sequence of up to 32 kilobases.
Similar primer selection programs have incorporated additional features for expanded capabilities. For example, the PrimOU primer selection program (available to the public from the Genome Center at University of I S Texas South West Medical Center, Dallas TX) is capable of choosing specific primers from megabase sequences and is thus useful for designing primers on a genome-wide scope. The Primer3 primer selection program (available to the public from the Whitehead lnstitute/M1T
Center for Genome Research, Cambridge MA) allows the user to input a "mispriming library," in which sequences to avoid as primer binding sites are user-specified. Primer3 is useful, in particular, for the selection of oligonucleotides for microarrays. (The source code for the latter two primer selection programs may also be obtained from their respective sources and modified to meet the user's specific needs.) The PrimeGen program (available to the public from the UK Human Genome Mapping Project Resource Centre, Cambridge UK) designs primers based on multiple sequence alignments, thereby allowing selection of primers that hybridize to either the most conserved or least conserved regions of aligned nucleic acid sequences. Hence, this program is useful for identification of both unique and conserved oligonucleotides and polynucleotide fragments. The oligonucleotides and polynucleotide fragments identified by any of the above selection methods are useful in hybridization technologies, for example, as PCR or sequencing primers, microarray elements, or specific probes to identify fully or partially complementary polynucleotides in a sample of nucleic acids. Methods of oligonucleotide selection are not limited to those described above.
A "recombinant nucleic acid" is a sequence that is not naturally occurring or has a sequence that is made by an artificial combination of two or more otherwise separated segments of sequence.
This artificial combination is often accomplished by chemical synthesis or, more commonly, by the artificial manipulation of isolated segments of nucleic acids, e.g., by genetic engineering techniques such as those described .in Sambrook, supra. The term recombinant includes nucleic acids that have been altered solely by addition, substitution, or deletion of a portion of the nucleic' acid. Frequently, a recombinant nucleic acid may include a nucleic acid sequence operably linked to a promoter sequence. Such a recombinant nucleic acid may be part of a vector that is used, for example, to transform a cell.
Alternatively, such recombinant nucleic acids may be part of a viral vector, e.g., based on a vaccinia virus, that could be use to vaccinate a mammal wherein the recombinant nucleic acid is expressed, inducing a protective immunological response in the mammal.
The term "sample" is used in its broadest sense. A sample suspected of containing nucleic acids encoding MTRP, or fragments thereof, or MTRP itself, may comprise a bodily fluid; an extract from a cell, chromosome, organelle, or membrane isolated from a cell; a cell;
genomic DNA, RNA, or cDNA, in solution or bound to a substrate; a tissue; a tissue print; etc.
The terms "specific binding" and "specifically binding" refer to that interaction between a protein or peptide and an agonist, an antibody, an antagonist, a small molecule, or any natural or synthetic binding composition. The interaction is dependent upon the presence of a particular structure of the protein, e.g., the antigenic determinant or epitope, recognized by the binding molecule. For example, if an antibody is specific for epitope "A," the presence of a polypeptide containing the epitope A, or the presence of free unlabeled A, in a reaction containing free labeled A
and the antibody will reduce the amount of labeled A that binds to the antibody.
The term "substantially purified" refers to nucleic acid or amino acid sequences that are removed from their natural environment and are isolated or separated, and are at least about 60% free, preferably about 75% free, and most preferably about 90% free from other components with which they are naturally associated.
A "substitution" refers to the replacement of one or more amino acids or nucleotides by different amino acids or nucleotides, respectively.
"Substrate" refers to any suitable rigid or semi-rigid support including membranes, filters, chips, slides, wafers, fibers, magnetic or nonmagnetic beads, gels, tubing, plates, polymers, microparticles and capillaries. The substrate can have a variety of surface forms, such as wells, trenches, pins, channels and pores, to which polynucleotides or polypeptides are bound.
"Transformation" describes a process by which exogenous DNA enters and changes a recipient cell. Transformation may occur under natural or artificial conditions according to various methods well known in the art, and may rely on any known method for the insertion of foreign nucleic acid sequences into a prokaryotic or eukaryotic host cell. The method for transformation is selected based on the type of host cell being transformed and may include, but is not limited to, viral infection, l9 electroporation, heat shock, lipofection, and particle bombardment. The term "transformed" cells includes stably transformed cells in which the inserted DNA is capable of replication either as an autonomously replicating plasmid or as part of the host chromosome, as well as transiently transformed cells which express the inserted DNA or RNA for limited periods of time.
A "variant" of a particular nucleic acid sequence is defined as a nucleic acid sequence having at least 40% sequence identity to the particular nucleic acid sequence over a certain length of one of the nucleic acid sequences using blastn with the "BLAST 2 Sequences" tool Version 2Ø9 (May-07-1999) set at default parameters. Such a pair of nucleic acids may show, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95% or at least 98% or greater sequence identity over a certain defined length. A variant may be described as, for example, an "allelic" (as defined above), "splice," "species," or "polymorphic"
variant. A splice variant may have significant identity to a reference molecule, but will generally have a greater or lesser number of polynucleotides due to alternate splicing of exons during mRNA processing. The corresponding polypeptide may possess additional functional domains or lack domains that are present in the IS reference molecule. Species variants are polynucleotide sequences that vary from one species to another. The resulting polypeptides generally will have significant amino acid identity relative to each other. A polymorphic variant is a variation in the polynucleotide sequence of a particular gene between individuals of a given species. Polymorphic variants also may encompass "single nucleotide polymorphisms" (SNPs) in which the polynucleotide sequence varies by one nucleotide base. The presence of SNPs may be indicative of, for example, a certain population, a disease state, or a propensity for a disease state.
A "variant" of a particular polypeptide sequence is defined as a polypeptide sequence having at least 40% sequence identity to the particular polypeptide sequence over a certain length of one of the polypeptide sequences using blastp with the "BLAST 2 Sequences" tool Version 2Ø9 (May-07-1999) set at default parameters. Such a pair of polypeptides may show, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 98% or greater sequence identity over a certain defined length of one of the polypeptides.
THE INVENTION
The invention is based on the discovery of new human membrane transport proteins (MTRP), the polynucleotides encoding MTRP, and the use of these compositions for the diagnosis, treatment, or prevention of membrane transport disorders; immune/inflammatory disorders;
and cell proliferative disorders including cancer.
Table I lists the Incyte clones used to assemble full length nucleotide sequences encoding MTRP. Columns 1 and 2 show the sequence identification numbers (SEQ ID NOs) of the polypeptide and nucleotide sequences, respectively. Column 3 shows the clone 1Ds of the Incyte clones in which nucleic acids encoding each MTRP were identified, and column 4 shows the cDNA
libraries from which these clones were isolated. Column 5 shows lncyte clones and their corresponding cDNA
libraries. Clones for which cDNA libraries are not indicated were derived from pooled cDNA
libraries. The Incyte clones in column S were used to assemble the consensus nucleotide sequence of each MTRP and are useful as fragments in hybridization technologies.
The columns of Table 2 show various properties of each of the polypeptides of the invention:
column 1 references the SEQ ID NO; column 2 shows the number of amino acid residues in each polypeptide; column 3 shows potential phosphorylation sites; column 4 shows potential glycosylation sites; column 5 shows the amino acid residues comprising signature sequences and motifs; column 6 shows homologous sequences as identified by BLAST analysis; and column 7 shows analytical methods and in some cases, searchable databases to which the analytical methods were applied. The methods of column 7 were used to characterize each polypeptide through sequence homology and protein motifs.
As shown in Figures 1 A, 1 B, 1 C, and 1 D, MTRP-3 has chemical and structural similarity with mouse fatty acid transport protein (FATP; GI 2612939; SEQ ID N0:35). In particular, MTRP-3 and FATP share 65% identity. As shown in Figures 2A, 2B, 2C, and 2D, MTRP-4 has chemical and structural similarity with Schistosoma mansoni ATP-binding cassette family protein, SMDR-1 (GI
425474; SEQ ID N0:36). In particular, MTRP-4 and SMDR-1 share 38% identity. As shown in Figures 3A, 3B, 3C, and 3D, MTRP-5 has chemical and structural similarity with rat sodium-dependent multivitamin transporter (SMVT; GI 3015617; SEQ ID N0:37). In particular, MTRP-5 and SMVT share 82% identity.
The columns of Table 3 show the tissue-specificity and diseases, disorders, or conditions associated with nucleotide sequences encoding MTRP. The first column of Table 3 lists the nucleotide SEQ ID NOs. Column 2 lists tissue categories which express MTRP as a fraction of total tissues expressing MTRP. Column 3 lists diseases, disorders, or conditions associated with those tissues expressing MTRP as a fraction of total tissues expressing MTRP. Column 4 lists the vectors used to subclone each cDNA library.
Of particular note are the expression patterns of SEQ ID N0:30 and SEQ ID
N0:31. SEQ ID
N0:30 is expressed in only five libraries, of which at least four (80%) are associated with cell proliferation and at least one (20%) with inflammation. Two (40%) ofthe five libraries are associated with cardiovascular tissue, and one each (20%) with gastrointestinal, nervous, and reproductive tissues. SEQ ID N0:31 is expressed in only four libraries, of which at least three (75%) are associated with cell proliferation and at least two (50%) with inflammation or the immune response.

Two (50%) of the four libraries are associated with hematopoietic/immune tissue, and one each (25%) with cardiovascular and reproductive tissues.
The following fragments of the nucleotide sequences encoding MTRP are useful, for example, in hybridization or amplification technologies to identify SEQ ID
N0:18-34 and to distinguish between SEQ ID N0:18-34 and related polynucieotide sequences. The useful fragments include the fragment of SEQ ID N0:18 from about nucleotide 110 to about nucleotide 154; the fragment of SEQ ID N0:19 from about nucleotide 759 to about nucleotide 839;
the fragment of SEQ
ID N0:20 from about nucleotide 1531 to about nucleotide 1578; the fragment of SEQ ID N0:21 from about nucleotide 538 to about nucleotide 597; the fragment of SEQ 1D N0:22 from about nucleotide 2241 to about nucleotide 2294; the fragment of SEQ ID N0:23 from about nucleotide 116 to about nucleotide 145; the fragment of SEQ ID N0:24 from about nucleotide 60 to about nucleotide 89; the fragment of SEQ ID N0:25 from about nucleotide 160 to about nucleotide 189;
the fragment of SEQ
ID N0:26 from about nucleotide 763 to about nucleotide 792; the fragment of SEQ ID N0:27 from about nucleotide 43 to about nucleotide 72; the fragment of SEQ ID N0:28 from about nucleotide 361 to about nucleotide 405; the fragment of SEQ ID N0:29 from about nucleotide 35 to about nucleotide 79; the fragment of SEQ ID N0:30 from about nucleotide 206 to about nucleotide 250; the fragment of SEQ ID N0:31 from about nucleotide 71 to about nucleotide 1 I5; the fragment of SEQ ID N0:32 from about nucleotide 161 to about nucleotide 205; the fragment of SEQ ID
N0:33 from about nucleotide 364 to about nucleotide 408; and the fragment of SEQ ID N0:34 from about nucleotide 18 to about nucleotide 62. The polypeptides encoded by the specified fragments of SEQ ID NO:20-30 and SEQ ID N0:32-34 are useful, for example, as immunogenic peptides.
The columns of Table 4 show descriptions of the tissues used to construct the cDNA libraries from which cDNA clones encoding MTRP were isolated. Column 1 references the nucleotide SEQ
1D NOs, column 2 shows the cDNA libraries from which these clones were isolated, and column 3 shows the tissue origins and other descriptive information relevant to the cDNA libraries in column 2.
The invention also encompasses MTRP variants. A preferred MTRP variant is one which has at least about 80%, or alternatively at least about 90%, or even at least about 95% amino acid sequence identity to the MTRP amino acid sequence, and which contains at least one functional or structural characteristic of MTRP.
The invention also encompasses polynucleotides which encode MTRP. In a particular embodiment, the invention encompasses a polynucleotide sequence comprising a sequence selected from the group consisting of SEQ ID N0:18-34, which encodes MTRP.
The invention also encompasses a variant of a polynucleotide sequence encoding MTRP. In particular, such a variant polynucleotide sequence will have at least about 75%, or alternatively at WO 00/26245 PC'T/US99/26048 least about 85%, or even at least about 95% polynucleotide sequence identity to the polynucleotide sequence encoding MTRP. A particular aspect of the invention encompasses a variant of a polynucleotide sequence comprising a sequence selected from the group consisting of SEQ ID
N0:18-34 which has at least about 75%, or alternatively at least about 85%, or even at least about 95% polynucleotide sequence identity to a nucleic acid sequence selected from the group consisting of SEQ ID N0:18-34. Any one of the polynucleotide variants described above can encode an amino acid sequence which contains at least one functional or structural characteristic of MTRP.
It will be appreciated by those skilled in the art that as a result of the degeneracy of the genetic code, a multitude of polynucleotide sequences encoding MTRP, some bearing minimal similarity to the polynucleotide sequences of any known and naturally occurring gene, may be produced. Thus, the invention contemplates each and every possible variation of polynucleotide sequence that could be made by selecting combinations based on possible codon choices. These combinations are made in accordance with the standard triplet genetic code as applied to the polynucleotide sequence of naturally occurring MTRP, and all such variations are to be considered as being specifically disclosed.
Although nucleotide sequences which encode MTRP and its variants are generally capable of hybridizing to the nucleotide sequence of the naturally occurring MTRP under appropriately selected conditions of stringency, it may be advantageous to produce nucleotide sequences encoding MTRP or its derivatives possessing a substantially different codon usage, e.g., inclusion of non-naturally occurring codons. Codons may be selected to increase the rate at which expression of the peptide occurs in a particular prokaryotic or eukaryotic host in accordance with the frequency with which particular codons are utilized by the host. Other reasons for substantially altering the nucleotide sequence encoding MTRP and its derivatives without altering the encoded amino acid sequences include the production of RNA transcripts having more desirable properties, such as a greater half life, than transcripts produced from the naturally occurring sequence.
The invention also encompasses production of DNA sequences which encode MTRP
and MTRP derivatives, or fragments thereof, entirely by synthetic chemistry. After production, the synthetic sequence may be inserted into any of the many available expression vectors and cell systems using reagents well known in the art. Moreover, synthetic chemistry may be used to introduce mutations into a sequence encoding MTRP or any fragment thereof.
Also encompassed by the invention are polynucleotide sequences that are capable of hybridizing to the claimed polynucleotide sequences, and, in particular, to those shown in SEQ ID
N0:18-34 and fragments thereof under various conditions of stringency. (See, e.g., Wahi, G.M. and S.L. Berger ( 1987) Methods Enzymol. 152:399-407; Kimmel, A.R. ( 1987) Methods Enzymol.

152:507-511.) Hybridization conditions, including annealing and wash conditions, are described in "Definitions."
Methods for DNA sequencing are well known in the art and may be used to practice any of the embodiments of the invention. The methods may employ such enzymes as the Klenow fragment of DNA polymerise 1, SEQUENASE (US Biochemical, Cleveland OH), Taq polymerise (Perkin-Elmer), thermostable T7 polymerise (Amersham Pharmacia Biotech, Piscataway NJ), or combinations of polymerises and proofreading exonucleases such as those found in the ELONGASE
amplification system (Life Technologies, Gaithersburg MD). Preferably, sequence preparation is automated with machines such as the MICROLAB 2200 liquid transfer system (Hamilton, Reno NV), PTC200 thermal cycler (MJ Research, Watertown MA) and ABI CATALYST 800 thermal cycler (Perkin-Elmer). Sequencing is then carried out using either the ABI 373 or 377 DNA sequencing system (Perkin-Elmer), the MEGABACE 1000 DNA sequencing system (Molecular Dynamics, Sunnyvale CA), or other systems known in the art. The resulting sequences are analyzed using a variety of algorithms which are well known in the art. (See, e.g., Ausubel, F.M. (1997) Short Protocols in Molecular Biolo~y, John Wiley & Sons, New York NY, unit 7.7;
Meyers, R.A. ( 1995) Molecular Biology and Biotechnolo>sy, Wiley VCH, New York NY, pp. 856-853.) The nucleic acid sequences encoding MTRP may be extended utilizing a partial nucleotide sequence and employing various PCR-based methods known in the art to detect upstream sequences, such as promoters and regulatory elements. For example, one method which may be employed, restriction-site PCR, uses universal and nested primers to amplify unknown sequence from genomic DNA within a cloning vector. (See, e.g., Sarkar, G. (1993) PCR Methods Applic.

2:318-322.) Another method, inverse PCR, uses primers that extend in divergent directions to amplify unknown sequence from a circularized template. The template is derived from restriction fragments comprising a known genomic locus and surrounding sequences. (See, e.g., Triglia, T. et al. (1988) Nucleic Acids Res. 16:8186.) A third method, capture PCR, involves PCR amplification of DNA
fragments adjacent to known sequences in human and yeast artificial chromosome DNA. (See, e.g., Lagerstrom, M. et al.
(1991 ) PCR Methods Applic. 1:111-119.) In this method, multiple restriction enzyme digestions and ligations may be used to insert an engineered double-stranded sequence into a region of unknown sequence before performing PCR. Other methods which may be used to retrieve unknown sequences are known in the art. (See, e.g., Parker, J.D. et al. ( 1991 ) Nucleic Acids Res. i 9:3055-3060).
Additionally, one may use PCR, nested primers, and PROMOTERFINDER libraries (Clontech, Palo Alto CA) to walk genomic DNA. This procedure avoids the need to screen libraries and is useful in finding intron/exon junctions. For all PCR-based methods, primers may be designed using commercially available software, such as OLIGO 4.06 Primer Analysis software (National Biosciences, Plymouth MN) or another appropriate program, to be about 22 to 30 nucleotides in length, to have a GC content of about 50% or more, and to anneal to the template at temperatures of about 68°C to 72°C.
When screening for full-length cDNAs, it is preferable to use libraries that have been size-selected to include larger cDNAs. In addition, random-primed libraries, which often include sequences containing the S' regions of genes, are preferable for situations in which an oligo d(T) Library does not yield a full-length cDNA. Genomic libraries may be useful for extension of sequence into S' non-transcribed regulatory regions.
Capillary electrophoresis systems which are commercially available may be used to analyze the size or confirm the nucleotide sequence of sequencing or PCR products. In particular, capillary sequencing may employ flowable polymers for electrophoretic separation, four different nucleotide specific, laser-stimulated fluorescent dyes, and a charge coupled device camera for detection of the emitted wavelengths. Output/light intensity may be converted to electrical signal using appropriate software {e.g., GENOTYPER and SEQUENCE NAVIGATOR, Perkin-Elmer), and the entire process I S from loading of samples to computer analysis and electronic data display may be computer controlled.
Capillary electrophoresis is especially preferable for sequencing small DNA
fragments which may be present in limited amounts in a particular sample.
In another embodiment of the invention, polynucleotide sequences or fragments thereof which encode MTRP may be cloned in recombinant DNA molecules that direct expression of MTRP, or fragments or functional equivalents thereof, in appropriate host cells. Due to the inherent degeneracy of the genetic code, other DNA sequences which encode substantially the same or a functionally equivalent amino acid sequence may be produced and used to express MTRP.
The nucleotide sequences of the present invention can be engineered using methods generally known in the art in order to alter MTRP-encoding sequences for a variety of purposes including, but not limited to, modification of the cloning, processing, and/or expression of the gene product. DNA
shuffling by random fragmentation and PCR reassembly of gene fragments and synthetic oligonucleotides may be used to engineer the nucleotide sequences. For example, oligonucleotide-mediated site-directed mutagenesis may be used to introduce mutations that create new restriction sites, alter glycosylation patterns, change codon preference, produce splice variants, and so forth.
In another embodiment, sequences encoding MTRP may be synthesized, in whole or in part, using chemical methods well known in the art. (See, e.g., Caruthers, M.H. et al. ( 1980) Nucleic Acids Symp. Ser. 7:215-223; and Horn, T. et al. ( 1980) Nucleic Acids Symp. Ser.
7:225-232.) Alternatively, MTRP itself or a fragment thereof may be synthesized using chemical methods. For example, peptide synthesis can be performed using various solid-phase techniques. (See, e.g., Roberge, J.Y. et al. (1995) Science 269:202-204.) Automated synthesis may be achieved using the ABI 431 A peptide synthesizer (Perkin-Elmer). Additionally, the amino acid sequence of MTRP, or any part thereof, may be altered during direct synthesis and/or combined with sequences from other proteins, or any part thereof, to produce a variant polypeptide.
The peptide may be substantially purified by preparative high performance liquid chromatography. (See, e.g., Chiez, R.M. and F.Z. Regnier (1990) Methods Enzymol. 182:392-421.) The composition of the synthetic peptides may be confirmed by amino acid analysis or by sequencing.
(See, e.g., Creighton, T. (1984) Proteins Structures and Molecular Proaerties, WH Freeman, New York NY.) In order to express a biologically active MTRP, the nucleotide sequences encoding MTRP or derivatives thereof may be inserted into an appropriate expression vector, i.e., a vector which contains the necessary elements for transcriptional and translational control of the inserted coding sequence in a suitable host. These elements include regulatory sequences, such as enhancers, constitutive and inducible promoters, and 5' and 3' untranslated regions in the vector and in polynucleotide sequences IS encoding MTRP. Such elements may vary in their strength and specificity.
Specific initiation signals may also be used to achieve more efficient translation of sequences encoding MTRP. Such signals include the ATG initiation codon and adjacent sequences, e.g. the Kozak sequence. In cases where sequences encoding MTRP and its initiation codon and upstream regulatory sequences are inserted into the appropriate expression vector, no additional transcriptional or translational control signals may be needed. However, in cases where only coding sequence, or a fragment thereof, is inserted, exogenous translational control signals including an in-frame ATG initiation codon should be provided by the vector. Exogenous translational elements and initiation codons may be of various origins, both natural and synthetic. The efficiency of expression may be enhanced by the inclusion of enhancers appropriate for the particular host cell system used. (See, e.g., Scharf, D. et al. ( 1994) Results Probl. Cell Differ. 20:125-162.) Methods which are well known to those skilled in the art may be used to construct expression vectors containing sequences encoding MTRP and appropriate transcriptional and translational control elements. These methods include in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. (See, e.g., Sambrook, J. et al. ( 1989) Molecular Cloning, A
Laboratory Manual, Cold Spring Harbor Press, Plainview NY, ch. 4, 8, and 16-17; Ausubel, F.M. et al. ( 1995) Current Protocols in Molecular Biolo~y, John Wiley & Sons, New York NY, ch. 9, 13, and 16.}
A variety of expression vector/host systems may be utilized to contain and express sequences encoding MTRP. These include, but are not limited to, microorganisms such as bacteria transformed with recombinant bacteriophage, plasmid, or cosmid DNA. expression vectors;
yeast transformed with yeast expression vectors; insect cell systems infected with viral expression vector's (e.g., baculovirus);
plant cell systems transformed with viral expression vectors (e.g., cauliflower mosaic virus, CaMV, or tobacco mosaic virus, TMV) or with bacterial expression vectors (e.g., Ti or pBR322 plasmids); or animal cell systems. The invention is not limited by the host cell employed.
In bacterial systems, a number of cloning and expression vectors may be selected depending upon the use intended for polynucleotide sequences encoding MTRP. For example, routine cloning, subcloning, and propagation of polynucleotide sequences encoding MTRP can be achieved using a multifunctional E. coli vector such as PBLUESCRIPT (Stratagene, La Jolla CA) or PSPORT1 plasmid (Life Technologies). Ligation of sequences encoding MTRP into the vector's multiple cloning site disrupts the lacZ gene, allowing a colorimetric screening procedure for identification of transformed bacteria containing recombinant molecules. 1n addition, these vectors may be useful for in vitro transcription, dideoxy sequencing, single strand rescue with helper phage, and creation of nested deletions in the cloned sequence. (See, e.g., Van Heeke, G. and S.M.
Schuster ( 1989) J. Biol.
Chem. 264:5503-5509.) When large quantities of MTRP are needed, e.g. for the production of antibodies, vectors which direct high level expression of MTRP may be used.
For example, vectors containing the strong, inducible TS or T7 bacteriophage promoter may be used.
Yeast expression systems may be used for production of MTRP. A number of vectors containing constitutive or inducible promoters, such as alpha factor, alcohol oxidase, and PGH
promoters, may be used in the yeast Saccharomyces cerevisiae or Pichia pastoris. In addition, such vectors direct either the secretion or intracellular retention of expressed proteins and enable integration of foreign sequences into the host genome for stable propagation.
(See, e.g:, Ausubel, 1995, su ra; Bitter, G.A. et al. ( 1987} Methods Enzymol. 153:516-544; and Scorer, C.A, et al. ( 1994) Bio/Technology 12:181-184.) Plant systems may also be used for expression of MTRP. Transcription of sequences encoding MTRP may be driven viral promoters, e.g., the 355 and 19S promoters of CaMV used alone or in combination with the omega leader sequence from TMV (Takamatsu, N.
(1987) EMBO J.
6:307-311 ). Alternatively, plant promoters such as the small subunit of RUBISCO or heat shock promoters may be used. (See, e.g., Coruzzi, G. et al. (1984) EMBO J. 3:1671-1680; Brogue, R. et al.
(1984) Science 224:838-843; and Winter, J. et al. (1991) Results Probl. Cell Differ. 17:85-105.) These constructs can be introduced into plant cells by direct DNA
transformation or pathogen-mediated transfection. (See, e.g., The McGraw Hill Yearbook of Science and Technoloev (1992) McGraw Hill, New York NY, pp. 191-196.) In mammalian cells, a number of viral-based expression systems may be utilized. In cases where an adenovirus is used as an expression vector, sequences encoding MTRP
may be ligated into an adenovirus transcription/translation complex consisting of the late promoter an'~ tripartite leader sequence. Insertion in a non-essential E1 or E3 region of the viral genome may be used to obtain infective virus which expresses MTRP in host cells. (See, e.g., Logan, J. and T. Shenk ( 1984) Proc.
Natl. Acad. Sci. USA 81:3655-3659.) In addition, transcription enhancers, such as the Rous sarcoma virus (RSV) enhancer, may be used to increase expression in mammalian host cells. SV40 or EBV-based vectors may also be used for high-level protein expression.
Human artificial chromosomes (HACs) may also be employed to deliver larger fragments of DNA than can be contained in and expressed from a plasmid. HACs of about 6 kb to 10 Mb are constructed and delivered via conventional delivery methods (liposomes, polycationic amino polymers, or vesicles) for therapeutic purposes. (See, e.g., Harrington, J.J.
et al. (1997) Nat. Genet.
15:345-355.) For long term production of recombinant proteins in mammalian systems, stable expression of MTRP in cell lines is preferred. For example, sequences encoding MTRP can be transformed into cell lines using expression vectors which may contain viral origins of replication and/or endogenous expression elements and a selectable marker gene on the same or on a separate vector. Following the introduction of the vector, cel Is may be allowed to grow for about 1 to 2 days in enriched media before being switched to selective media. The purpose of the selectable marker is to confer resistance to a selective agent, and its presence allows growth and recovery of cells which successfully express the introduced sequences. Resistant clones of stably transformed cells may be propagated using tissue culture techniques appropriate to the cell type.
Any number of selection systems may be used to recover transformed cell lines:
These include, but are not limited to, the herpes simplex virus thymidine kinase and adenine phosphoribosyltransferase genes, for use in tk- and apr~ cells, respectively.
(See, e.g., Wigler, M. et al. (1977) Cell 11:223-232; Lowy, I. et al. (1980) Cell 22:817-823.) Also, antimetabolite, antibiotic, or herbicide resistance can be used as the basis for selection. For example, dhfr confers resistance to methotrexate; neo confers resistance to the aminoglycosides neomycin and G-418; and als and pat confer resistance to chlorsulfuron and phosphinotricin acetyltransferase, respectively. (See, e.g., Wigler, M. et al: (1980) Proc. Natl. Acad. Sci. USA 77:3567-3570; Colbere-Garapin, F. et al. (1981) J. Mol. Biol. 150:1-14.) Additional selectable genes have been described, e.g., trpB and hisD, which alter cellular requirements for metabolites. (See, e.g., Hartman, S.C. and R.C. Mulligan ( 1988) Proc.
Natl. Acad. Sci. USA 85:8047-8051.) Visible markers, e.g., anthocyanins, green fluorescent proteins (GFP; Clontech), b glucuronidase and its substrate (3-glucuronide, or iuciferase and its substrate luciferin may be used. These markers can be used not only to identify transformants, but also to quantify the amount of transient or stable protein expression attributable to a specific vector system.
(See, e.g., Rhodes, C.A. (1995) Methods Mol. Biol. 55:121-131.) Although the presence/absence of marker gene expression suggests that the gene of interest is also present, the presence and expression of the gene may need to be confirmed. For example, if the sequence encoding MTRP is inserted within a marker gene sequence, transformed cells containing sequences encoding MTRP can be identified by the absence of marker gene function. Alternatively, a marker gene can be placed in tandem with a sequence encoding MTRP under the control of a single promoter. Expression of the marker gene in response to induction or selection usually indicates expression of the tandem gene as well.
In general, host cells that contain the nucleic acid sequence encoding MTRP
and that express MTRP may be identified by a variety of procedures known to those of skill in the art. These procedures include, but are not limited to, DNA-DNA or DNA-RNA hybridizations, PCR
amplification, and protein bioassay or immunoassay techniques which include membrane, solution, or chip based technologies for the detection and/or quantification of nucleic acid or protein sequences.
lmmunological methods for detecting and measuring the expression of MTRP using either specific polyclonal or monoclonal antibodies are known in the art. Examples of such techniques include enzyme-linked immunosorbent assays (ELISAs), radioimmunoassays (RIAs), and fluorescence activated cell sorting (FACS). A two-site, monoclonal-based immunoassay utilizing monoclonal antibodies reactive to two non-interfering epitopes on MTRP is preferred, but a competitive binding assay may be employed. These and other assays are well known in the art. (See, e.g., Hampton, R. et al. ( 1990) Serological Methods, a Laboratory Manual, APS
Press, St. Paul MN, Sect. 1V; Coligan, J.E. et al. (1997) Current Protocols in Immunoloay, Greene Pub. Associates and Wiley-Interscience, New York NY; and Pound, J.D. (1998) lmmunochemical Protocols, Humana Press, Totowa NJ.) A wide variety of labels and conjugation techniques are known by those skilled in the art and may be used in various nucleic acid and amino acid assays. Means for producing labeled hybridization or PCR probes for detecting sequences related to polynucleotides encoding MTRP
include oligolabeling, nick translation, end-labeling, or PCR amplification using a labeled nucleotide.
Alternatively, the sequences encoding MTRP, or any fragments thereof, may be cloned into a vector for the production of an mRNA probe. Such vectors are known in the art, are commercially availabie, and may be used to synthesize RNA probes in vitro by addition of an appropriate RNA polymerase such as T7, T3, or SP6 and labeled nucleotides. These procedures may be conducted using a variety of commercially available kits, such as those provided by Amersham Pharmacia Biotech, Promega (Madison WI), and US Biochemical. Suitable reporter molecules or labels which may be used for ease of detection include radionuclides, enzymes, fluorescent, chemiluminescent, or chromogenic agents, as well as substrates, cofactors, inhibitors, magnetic particles, and the like.
Host cells transformed with nucleotide sequences encoding MTRP may be cultured under conditions suitable for the expression and recovery of the protein from cell culture. The protein S produced by a transformed cell may be secreted or retained intracellularly depending on the sequence and/or the vector used. As will be understood by those of skill in the art, expression vectors containing polynucleotides which encode MTRP may be designed to contain signal sequences which direct secretion of MTRP through a prokaryotic or eukaryotic cell membrane.
In addition, a host cell strain may be chosen for its ability to modulate expression of the inserted sequences or to process the expressed protein in the desired fashion.
Such modifications of the polypeptide include, but are not limited to, acetylation, carboxylation, glycosylation, phosphorylation, lipidation, and acylation. Post-translational processing which cleaves a "prepro" or "pro" form of the protein may also be used to specify protein targeting, folding, and/or activity.
Different host cells which have specific cellular machinery and characteristic mechanisms for post-translational activities (e.g., CHO, HeLa, MDCK, HEK293, and WI38) are available from the American Type Culture Collection (ATCC, Manassas VA) and may be chosen to ensure the correct modification and processing of the foreign protein.
In another embodiment of the invention, natural, modified, or recombinant nucleic acid sequences encoding MTRP may be ligated to a heterologous sequence resulting in translation of a fusion protein in any of the aforementioned host systems. For example, a chimeric MTRP protein containing a heterologous moiety that can be recognized by a commercially available antibody may facilitate the screening of peptide libraries for inhibitors of MTRP activity.
Heterologous protein and peptide moieties may also facilitate purification of fusion proteins using commercially available affinity matrices. Such moieties include, but are not limited to, glutathione S-transferase (GST), maltose binding protein (MBP), thioredoxin (Trx), calmodulin binding peptide (CBP), 6-His, FLAG, c-myc, and hemagglutinin (HA). GST, MBP, Trx, CBP, and 6-His enable purification of their cognate fusion proteins on immobilized glutathione, .maltose, phenylarsine oxide, calmodulin, and metal-chelate resins, respectively. FLAG, c-myc, and hemagglutinin (HA) enable immunoaffinity purification of fusion proteins using commercially available monoclonal and polyclonal antibodies that specifically recognize these epitope tags. A fusion protein may also be engineered to contain a proteolytic cleavage site located between the MTRP encoding sequence and the heterologous protein sequence, so that MTRP may be cleaved away from the heterologous moiety following purification.
Methods for fusion protein expression and purification are discussed in Ausubel ( 1995, suara, ch. 10).
A variety of commercially available kits may also be used to facilitate expression and purification of fusion proteins.
In a further embodiment of the invention, synthesis of radiolabeled MTR~' may be achieved in vitro using the TNT rabbit reticulocyte lysate or wheat germ extract system (Promega). These systems couple transcription and translation of protein-coding sequences operably associated with the T7, T3, or SP6 promoters. Translation takes place in the presence of a radiolabeled amino acid precursor, for example, 'SS-methionine.
Fragments of MTRP may be produced not only by recombinant means, but also by direct peptide synthesis using solid-phase techniques. (See, e.g., Creighton, supra, pp. 55-60.) Protein synthesis may be performed by manual techniques or by automation. Automated synthesis may be achieved, for example, using the ABI 431 A peptide synthesizer (Perkin-Elmer).
Various fragments of MTRP may be synthesized separately and then combined to produce the ful) length molecule.
THERAPEUTICS
Chemical and structural similarity, e.g., in the context of sequences and motifs, exists between regions of MTRP and membrane transport proteins, including amino acid transporters, ABC
transporters, nucleotide-sugar transporters, transmembrane carrier proteins, and ATP-dependent transporter proteins. In addition, the expression of MTRP is closely associated with nervous, reproductive, and gastrointestinal tissues; cancer and other cell proliferative conditions; and with inflammation and the immune response. Therefore, MTRP appears to play a role in membrane transport disorders; immune/inflammatory disorders; and cell proliferative disorders including cancer.
In the treatment of disorders associated with increased MTRP expression or activity, it is desirable to decrease the expression or activity of MTRP. In the treatment of disorders associated with decreased MTRP expression or activity, it is desirable to increase the expression or activity of MTRP.
Therefore, in one embodiment, MTRP or a fragment or derivative thereof may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of MTRP. Examples of such disorders include, but are not limited to, a membrane transport disorder such as cystinuria, dibasicaminoaciduria, hypercystinuria, lysinuria, hartnup disease, tryptophan malabsorption, methionine malabsorption, histidinuria, iminoglycinuria, dicarboxylicaminoaciduria, cystinosis, renal glycosuria, glucose-galactose malabsorption, familial hypercholesterolemia, hypouricemia, familial hypophophatemic rickets, congenital chloridorrhea, cystic fibrosis, familial goiter, distal renal tubular acidosis, Menkes' disease, lethal diarrhea, nephrogenic diabetes insipidus, juvenile pernicious anemia, folate malabsorption, adrenoleukodystrophy, hereditary myoglobinuria, Zellweger syndrome, hyperinsulinemic hypoglycemia, akinesia, amyotrophic lateral sclerosis, ataxia telangiectasia, cystic fibrosis, Becker's muscular dystrophy, Bell's palsy, Charcot-Marie Tooth disease, diabetes mellitus, diabetes insipidus, diabetic neuropathy, Duchenne muscular dystrophy, hyperkalemic periodic paralysis, normokalemic periodic paralysis, Parkinson's disease, malignant hyperthermia, multidrug resistance, myasthenia gravis, myotonic dystrophy, catatonia, tardive dyskinesia, dystonias, peripheral neuropathy, cerebral neoplasms, and prostate cancer; a cardiac disorder associated with transport such as angina, bradyarrythmia, tachyarrythmia, hypertension, Long QT syndrome, myocarditis, cardiomyopathy, nemaline myopathy, centronuclear myopathy, lipid myopathy, mitochondrial myopathy, thyrotoxic myopathy, ethanol myopathy, dermatomyositis, inclusion body myositis, infectious myositis, and polymyositis; a neurological disorder associated with transport such as Alzheimer's disease, amnesia, bipolar disorder, dementia, depression, epilepsy, Tourette's disorder, paranoid psychoses, and schizophrenia; and an other disorder associated with transport such as neurofibromatosis, postherpetic neuralgia, trigeminal neuropathy, sarcoidosis, sickle cell anemia, Wilson's disease, cataracts, infertility, pulmonary artery stenosis, sensorineural autosomal deafness, hyperglycemia, hypoglycemia, Grave's disease, goiter, Cushing's disease, Addison's disease, glucose-galactose malabsorption syndrome, and hypercholesterolemia; an immune/inflammatory disorder such as acquired immunodeficiency syndrome (AIDS), Addison's disease, adult respiratory distress syndrome, allergies, ankylosing spondylitis, amyloidosis, anemia, asthma, atherosclerosis, autoimmune hemolytic anemia, autoimmune thyroiditis, autoimmune polyenodocrinopathy-candidiasis-ectodermal dystrophy (APECED), bronchitis, chotecystitis, contact dermatitis, Crohn's disease, atopic dermatitis, dermatomyositis, diabetes mellitus, emphysema, episodic lymphopenia with lymphocytotoxins, erythroblastosis fetalis, erythema nodosum, atrophic gastritis, glomerulonephritis, Goodpasture's syndrome, gout, Graves' disease, Hashimoto's thyroiditis, hypereosinophilia, irritable bowel syndrome, multiple sclerosis, myasthenia gravis, myocardial or pericardial inflammation, osteoarthritis, osteoporosis, pancreatitis, polymyositis, psoriasis, Reiter's syndrome, rheumatoid arthritis, scferoderma, Sjogren's syndrome, systemic anaphylaxis, systemic lupus erythematosus, systemic sclerosis, thrombocytopenic purpura, ulcerative colitis, uveitis, Werner syndrome, complications of cancer, hemodialysis, and extracorporeal circulation; a viral, bacterial, fungal, parasitic, protozoal, or helminthic infection; and trauma; a cell proliferative disorder such as actinic keratosis, arteriosclerosis, atherosclerosis, bursitis, cirrhosis, hepatitis, mixed connective tissue disease (MCTD), myelofibrosis, paroxysmal nocturnal hemoglobinuria, polycythemia vera, psoriasis, primary thrombocythemia; and a cancer including adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, teratocarcinoma; and, in particular, a cancer of the adrenal gland, bladder, bone, bone marrow, brain, breast, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, penis, prostate, salivary glands, skin, spleen, testis, thymus, thyroid, and uterus.

In another embodiment, a vector capable of expressing MTRP or a fragment or derivative thereof may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of MTRP including, but not limited to, those described above.
In a further embodiment, a pharmaceutical composition comprising a substantially purified MTRP in conjunction with a suitable pharmaceutical carrier may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of MTRP
including, but not limited to, those provided above.
In still another embodiment, an agonist which modulates the activity of MTRP
may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of MTRP including, but not limited to, those listed above.
In a further embodiment, an antagonist of MTRP may be administered to a subject to treat or prevent a disorder associated with increased expression or activity of MTRP.
Examples of such disorders include, but are not limited to, those membrane transport disorders;
immune/inflammatory disorders; and cell proiiferative disorders including cancer described above.
In one aspect, an antibody which specifically binds MTRP may be used directly as an antagonist or indirectly as a targeting or delivery mechanism for bringing a pharmaceutical agent to cells or tissues which express MTRP.
In an additional embodiment, a vector expressing the complement of the polynucleotide encoding MTRP may be administered to a subject to treat or prevent a disorder associated with increased expression or activity of MTRP including, but not limited to, those described above.
In other embodiments, any of the proteins, antagonists, antibodies, agonists, complementary sequences, or vectors of the invention may be administered in combination with other appropriate therapeutic agents. Selection of the appropriate agents for use in combination therapy may be made by one of ordinary skill in the art, according to conventional pharmaceutical principles. The combination of therapeutic agents may act synergistically to effect the treatment or prevention of the various disorders described above. Using this approach, one may be able to achieve therapeutic efficacy with lower dosages of each agent, thus reducing the potential for adverse side effects.
An antagonist of MTRP may be produced using methods which are generally known in the art. In particular, purified MTRP may be used to produce antibodies or to screen libraries of pharmaceutical agents to identify those which specifically bind MTRP.
Antibodies to MTRP may also be generated using methods that are well known in the art. Such antibodies may include, but are not limited to, polyclonal, monoclonal, chimeric, and single chain antibodies, Fab fragments, and fragments produced by a Fab expression library. Neutralizing antibodies (i.e., those which inhibit dimer formation) are generally preferred for therapeutic use.

For the production of antibodies, various hosts including goats, rabbits, rats, mice, humans, and others may be immunized by injection with MTRP or with any fragment or oligopeptide thereof which has immunogenic properties. Depending on the host species, various adjuvants may be used to increase immunological response. Such adjuvants include, but are not limited to, Freund's, mineral gels such as aluminum hydroxide, and surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, KLH, and dinitrophenol. Among adjuvants used in humans, BCG (bacilli Calmette-Guerin) and Cor_vnebacterium ~arvum are especially preferable.
It is preferred that the oligopeptides, peptides, or fragments used to induce antibodies to MTRP have an amino acid sequence consisting of at least about 5 amino acids, and generally will consist of at least about 10 amino acids. It is also preferable that these oligopeptides, peptides, or fragments are identical to a portion of the amino acid sequence of the natural protein and contain the entire amino acid sequence of a small, naturally occurring molecule. Short stretches of MTRP amino acids may be fused with those of another protein, such as KLH, and antibodies to the chimeric molecule may be produced.
Monoclonal antibodies to MTRP may be prepared using any technique which provides for the production of antibody molecules by continuous cell lines in culture. These include, but are not limited to, the hybridoma technique, the human B-cell hybridoma technique, and the EBV-hybridoma technique. (See, e.g., Kohler, G. et al. (1975) Nature 256:495-497; Kozbor, D.
et al. (1985) J.
Immunol. Methods 81:31-42; Cote, R.J. et al. ( 1983) Proc. Natl. Acad. Sci.
USA 80:2026-2030; and Cole, S.P. et al. (1984) Mol. Cell Biol. 62:109-120.) In addition, techniques developed for the production of "chimeric antibodies,"
such as the splicing of mouse antibody genes to human antibody genes to obtain a molecule with appropriate antigen specificity and biological activity, can be used. (See, e.g., Morrison, S.L. et al. (1984) Proc.
Natl. Acad. Sci. USA 81:6851-6855; Neuberger, M.S. et al. (1984) Nature 312:604-608; and Takeda, S. et al. ( 1985) Nature 314:452-454.) Alternatively, techniques described for the production of single chain antibodies may be adapted, using methods known in the art, to produce MTRP-specific single chain antibodies. Antibodies with related specificity, but of distinct idiotypic composition, may be generated by chain shuffling from random combinatorial immunoglobulin libraries. (See, e.g., Burton, D.R. ( 1991 ) Proc. Natl. Acad. Sci. USA 88:10134-10137.) Antibodies may also be produced by inducing in vivo production in the lymphocyte population or by screening immunoglobulin libraries or panels of highly specific binding reagents as disclosed in the literature. (See, e.g., Orlandi, R. et al. (1989) Proc. Natl.
Acad. Sci. USA
86:3833-3837; Winter, G. et al. (1991) Nature 349:293-299.) Antibody fragments which contain specific binding sites for MTRP may also be generated.

For example, such fragments include, but are not limited to, F(ab'), fragments produced by pepsin digestion of the antibody molecule and Fab fragments generated by reducing the disulfide bridges of the F(ab')2 fragments. Alternatively, Fab expression libraries may be constructed to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity.
(See, e.g., Huse, W.D.
et al. (1989) Science 246:1275-1281.) Various immunoassays may be used for screening to identify antibodies having the desired specificity. Numerous protocols for competitive binding or immunoradiometric assays using either polyclonal or monoclonal antibodies with established specificities are well known in the art. Such immunoassays typically involve the measurement of complex formation between MTRP and its specific antibody. A two-site, monoclonal-based immunoassay utilizing monoclonal antibodies reactive to two non-interfering MTRP epitopes is generally used, but a competitive binding assay may also be employed (Pound, supra).
Various methods such as Scatchard analysis in conjunction with radioimmunoassay techniques may be used to assess the affinity of antibodies for MTRP. Affinity is expressed as an I S association constant, K" which is defined as the molar concentration of MTRP-antibody complex divided by the molar concentrations of free antigen and free antibody under equilibrium conditions.
The K, determined for a preparation of polyclonal antibodies, which are heterogeneous in their affinities for multiple MTRP epitopes, represents the average affinity, or avidity, of the antibodies for MTRP. The K8 determined for a preparation of monoclonal antibodies, which are monospecific for a particular MTRP epitope, represents a true measure of affinity. High-affinity antibody preparations with Ka ranging from about 10''to 10''- L/mole are preferred for use in immunoassays in which the MTRP-antibody complex must withstand rigorous manipulations. Low-affinity antibody preparations with Ke ranging from about 106 to 10' L/mole are preferred for use in immunopurification and similar procedures which ultimately require dissociation of MTRP, preferably in active form, from the antibody (Catty, D. (1988) Antibodies, Volume I: A Practical Approach, IRL
Press, Washington, DC;
Liddell, J.E. and Cryer, A. (1991} A Practical Guide to Monoclonal Antibodies, John Wiley & Sons, New York NY).
The titer and avidity of polyclonal antibody preparations may be further evaluated to determine the quality and suitability of such preparations for certain downstream applications. For example, a polyclonal antibody preparation containing at least 1-2 mg specific antibody/ml, preferably 5-10 mg specific antibody/ml, is generally employed in procedures requiring precipitation of MTRP-antibody complexes. Procedures for evaluating antibody specificity, titer, and avidity, and guidelines for antibody quality and usage in various applications, are generally available. (See, e.g., Catty, supra, and Coligan et al. supra.) In another embodiment of the invention, the polynucleotides encoding MTRP, or any fragment or complement thereof, may be used for therapeutic purposes. In one aspect, the complement of the polynucleotide encoding MTRP may be used in situations in which it would be desirable to block the transcription of the mRNA. In particular, cells may be transformed with sequences complementary to polynucleotides encoding MTRP. Thus, complementary molecules or fragments may be used to modulate MTRP activity, or to achieve regulation of gene function. Such technology is now well known in the art, and sense or antisense oligonucleotides or larger fragments can be designed from various locations along the coding or control regions of sequences encoding MTRP.
Expression vectors derived from retroviruses, adenoviruses, or herpes or vaccinia viruses, or from various bacterial plasmids, may be used for delivery of nucleotide sequences to the targeted organ, tissue, or cell population. Methods which are well known to those skilled in the art can be used to construct vectors to express nucleic acid sequences complementary to the polynucleotides encoding MTRP. (See, e.g., Sambrook, su ra; Ausubel, 1995, supra.) Genes encoding MTRP can be turned off by transforming a cell or tissue with expression vectors which express high levels of a polynucleotide, or fragment thereof, encoding MTRP. Such constructs may be used to introduce untranslatable sense or antisense sequences into a cell. Even in the absence of integration into the DNA, such vectors may continue to transcribe RNA molecules until they are disabled by endogenous nucleases. Transient expression may last for a month or more with a non-replicating vector, and may last even longer if appropriate replication elements are part of the vector system.
As mentioned above, modifications of gene expression can be obtained by designing complementary sequences or antisense molecules (DNA, RNA, or PNA) to the control, 5', or regulatory regions of the gene encoding MTRP. Oligonucleotides derived from the transcription initiation site, e.g., between about positions -10 and +10 from the start site, may be employed.
Similarly, inhibition can be achieved using triple helix base-pairing methodology. Triple helix pairing is useful because it causes inhibition of the ability of the double helix to open sufficiently for the binding of polymerases, transcription factors, or regulatory molecules. Recent therapeutic advances using triplex DNA have been described in the literature. (See, e.g., Gee, J.E.
et al. (1994) in Huber, B.E. and B.I. Carr, Molecular and Immunolo i; ~ c Approaches, Futura Publishing, Mt. Kisco NY, pp.
163-177.) A complementary sequence or antisense molecule may also be designed to block translation of mRNA by preventing the transcript from binding to ribosomes.
Ribozymes, enzymatic RNA molecules, may also be used to catalyze the specific cleavage of RNA. The mechanism of ribozyme action involves sequence-specific hybridization of the ribozyme molecule to complementary target RNA, followed by endor~ucleolytic cleavage.
For example, engineered hammerhead motif ribozyme molecules may specifically and efficiently catalyze endonucleolytic cleavage of sequences encoding MTRP.
Specific ribozyme cleavage sites within any potential RNA target are initially identified by scanning the target molecule for ribozyme cleavage sites, including the following sequences: GUA, GUU, and GUC. Once identified, short RNA sequences of between 15 and 20 ribonucleotides, corresponding to the region of the target gene containing the cleavage site, may be evaluated for secondary structural features which may render the oligonucleotide inoperable.
The suitability of candidate targets may also be evaluated by testing accessibility to hybridization with complementary oligonucleotides using ribonuclease protection assays.
Complementary ribonucleic acid molecules and ribozymes of the invention may be prepared by any method known in the art for the synthesis of nucleic acid molecules.
These include techniques for chemically synthesizing oligonucleotides such as solid phase phosphoramidite chemical synthesis.
Alternatively, RNA molecules may be generated by in vitro and in vivo transcription of DNA
sequences encoding MTRP. Such DNA sequences may be incorporated into a wide variety of vectors with suitable RNA polymerase promoters such as T7 or SP6. Alternatively, these cDNA constructs that synthesize complementary RNA, constitutively or inducibly, can be introduced into cell lines, cel Is, or tissues.
RNA molecules may be modified to increase intracellular stability and half life. Possible modifications include, but are not limited to, the addition of flanking sequences at the 5' and/or 3' ends of the molecule, or the use of phosphorothioate or 2' O-methyl rather than phosphodiesterase linkages within the backbone of the molecule. This concept is inherent in the production of PNAs and can be extended in all of these molecules by the inclusion of nontraditional bases such as inosine, queosine, and wybutosine, as well as acetyl-, methyl-, thio-, and similarly modified forms of adenine, cytidine, guanine, thymine, and uridine which are not as easily recognized by endogenous endonucleases.
Many methods for introducing vectors into cells or tissues are available and equally suitable for use in vivo, in vitro, and ex vivo. For ex vivo therapy, vectors may be introduced into stem cells taken from the patient and clonally propagated for autologous transplant back into that same patient.
Delivery by transfection, by liposome injections, or by polycationic amino polymers may be achieved using methods which are well known in the art. (See, e.g., Goldman, C.K. et al. (1997) Nat.
Biotechnol. 15:462-466.) Any of the therapeutic methods described above may be applied to any subject in need of such therapy, including, for example, mammals such as humans, dogs, cats, cows, horses, rabbits, and monkeys.

An additional embodiment of the invention relates,to the administration of a pharmaceutical or sterile composition, in conjunction with a pharmaceutically acceptable carrier,'for any of the therapeutic effects discussed above. Such pharmaceutical compositions may consist of MTRP, antibodies to MTRP, and mimetics, agonists, antagonists, or inhibitors of MTRP. The compositions may be administered alone or in combination with at least one other agent, such as a stabilizing compound, which may be administered in any sterile, biocompatible pharmaceutical carrier including, but not limited to, saline, buffered saline, dextrose, and water. The compositions may be administered to a patient alone, or in combination with other agents, drugs, or hormones.
The pharmaceutical compositions utilized in this invention may be administered by any number of routes including, but not limited to, oral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, intraventricular, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, or rectal means.
In addition to the active ingredients, these pharmaceutical compositions may contain suitable pharmaceutically-acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically.
Further details on techniques for formulation and administration may be found in the latest edition of Remin tg on's Pharmaceutical Sciences (Maack Publishing, Easton PA).
Pharmaceutical compositions for oral administration can be formulated using pharmaceutically acceptable carriers well known in the art in dosages suitable for oral administration.
Such carriers enable the pharmaceutical compositions to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, and the like, for ingestion by the patient.
Pharmaceutical preparations for oral use can be obtained through combining active compounds with solid excipient and processing the resultant mixture of granules (optionally, after grinding) to obtain tablets or dragee cores. Suitable auxiliaries can be added, if desired. Suitable excipients include carbohydrate or protein fillers, such as sugars, including lactose, sucrose, mannitol, and sorbitol; starch from corn, wheat, rice, potato, or other plants;
cellulose, such as methyl cellulose, hydroxypropylmethyl-cellulose, or sodium carboxymethylcellulose; gums, including arabic and tragacanth; and proteins, such as gelatin and collagen. if desired, disintegrating or solubilizing agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, and alginic acid or a salt thereof, such as sodium alginate.
Dragee cores may be used in conjunction with suitable coatings, such as concentrated sugar solutions, which may also contain gum arabic, talc, polyvinylpyrrolidone, carbopo) gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures.
Dyestuffs or pigments may be added to the tablets or dragee coatings for product identification or to characterize the quantity of active compound, i.e., dosage. .
Pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a coating, such as glycerol or sorbitol.
Push-fit capsules can contain active ingredients mixed with fillers or binders, such as lactose or starches, lubricants, such as talc or magnesium stearate, and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid, or liquid polyethylene glycol with or without stabilizers.
Pharmaceutical formulations suitable for parenteral administration may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hanks' solution, Ringer's solution, or physiologically buffered saline. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Additionally, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oieate, triglycerides, or liposomes. Non-lipid polycationic IS amino polymers may also be used for delivery. Optionally, the suspension may also contain suitable stabilizers or agents to increase the solubility of the compounds and allow for the preparation of highly concentrated solutions.
For topical or nasal administration, penetrants appropriate to the particular barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.
The pharmaceutical compositions of the present invention may be manufactured in a manner that is known in the art, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilizing processes.
The pharmaceutical composition may be provided as a salt and can be formed with many acids, including but not limited to, hydrochloric, sulfuric, acetic, lactic, tartaric, malic, and succinic acids. Salts tend to be more soluble in aqueous or other protonic solvents than are the corresponding free base forms. In other cases, the preparation may be a lyophilized powder which may contain any or all ofthe following: I mM to 50 mM histidine, 0.1% to 2% sucrose, and 2% to 7% mannitol, at a pH range of 4.5 to 5.5, that is combined with buffer prior to use.
After pharmaceutical compositions have been prepared, they can be placed in an appropriate container and labeled for treatment of an indicated condition. For administration of MTRP, such labeling would include amount, frequency, and method of administration.
Pharmaceutical compositions suitable for use in the invention include compositions wherein the active ingredients are contained in an effective amount to achieve the intended purpose. The determination of an effective dose is well within the capability of those skilled in the art.

For any compound, the therapeutically effective dose can be estimated initially either in cell culture assays, e.g., of neoplastic cells, or in animal models such as mice, rats, rabbits, dogs, or pigs.
An animal model may also be used to determine the appropriate concentration range and route of administration. Such information can then be used to determine useful doses and routes for administration in humans.
A therapeutically effective dose refers to that amount of active ingredient, for example MTRP
or fragments thereof, antibodies of MTRP, and agonists, antagonists or inhibitors of MTRP, which ameliorates the symptoms or condition. Therapeutic efficacy and toxicity may be determined by standard pharmaceutical procedures in cell cultures or with experimental animals, such as by calculating the EDT (the dose therapeutically effective in 50% of the population) or LDS° (the dose lethal to 50% of the population) statistics. The dose ratio of toxic to therapeutic effects is the therapeutic index, which can be expressed as the LDS°/EDs°
ratio. Pharmaceutical compositions which exhibit large therapeutic indices are preferred. The data obtained from cell culture assays and animal studies are used to formulate a range of dosage for human use. The dosage contained in such compositions is preferably within a range of circulating concentrations that includes the EDS° with little or no toxicity. The dosage varies within this range depending upon the dosage form employed, the sensitivity of the patient, and the route of administration.
The exact dosage will be determined by the practitioner, in light of factors related to the subject requiring treatment. Dosage and administration are adjusted to provide sufficient levels of the active moiety or to maintain the desired effect. Factors which may be taken into account include the severity of the disease state, the general health of the subject, the age, weight, and gender of the subject, time and frequency of administration, drug combination(s), reaction sensitivities, and response to therapy. Long-acting pharmaceutical compositions may be administered every 3 to 4 days, every week, or biweekly depending on the half life and clearance rate of the particular formulation.
Normal dosage amounts may vary from about 0.1 ~g to 100,000 ~cg, up to a total dose of about 1 gram, depending upon the route of administration. Guidance as to particular dosages and methods of delivery is provided in the literature and generally available to practitioners in the art.
Those skilled in the art will employ different formulations for nucleotides than for proteins or their inhibitors. Similarly, delivery of polynucleotides or polypeptides will be specific to particular cells, conditions, locations, etc.
DIAGNOSTICS
In another embodiment, antibodies which specifically bind MTRP may be used for the diagnosis of disorders characterized by expression of MTRP, or in assays to monitor patients being treated with MTRP or agonists, antagonists, or inhibitors of MTRP. Antibodies useful for diagnostic purposes may be prepared in the same manner as described above for therapeutics: Diagnostic assays for MTRP include methods which utilize the antibody and a label to detect MTRP
in human body fluids or in extracts of cells or tissues. The antibodies may be used with or without modification, and may be labeled by covalent or non-covalent attachment of a reporter molecule.
A wide variety of reporter molecules, several of which are described above, are known in the art and may be used.
A variety of protocols for measuring MTRP, including ELISAs, RIAs, and FACS, are known in the art and provide a basis for diagnosing altered or abnormal levels of MTRP expression. Normal or standard values for MTRP expression are established by combining body fluids or cell extracts taken from normal mammalian subjects, for example, human subjects, with antibody to MTRP under conditions suitable for complex formation. The amount of standard complex formation may be quantitated by various methods, such as photometric means. Quantities of MTRP
expressed in subject, control, and disease samples from biopsied tissues are compared with the standard values.
Deviation between standard and subject values establishes the parameters for diagnosing disease.
In another embodiment of the invention, the polynucleotides encoding MTRP may be used for diagnostic purposes. The polynucleotides which may be used include oligonucleotide sequences, complementary RNA and DNA molecules, and PNAs. The polynucleotides may be used to detect and quantify gene expression in biopsied tissues in which expression of MTRP
may be correlated with disease. The diagnostic assay may be used to determine absence, presence, and excess expression of MTRP, and to monitor regulation of MTRP levels during therapeutic intervention.
In one aspect, hybridization with PCR probes which are capable of detecting polynucleotide sequences, including genomic sequences, encoding MTRP or closely related molecules may be used to identify nucleic acid sequences which encode MTRP. The specificity of the probe, whether it is made from a highly specific region, e.g., the 5' regulatory region, or from a Less specific region, e.g., a conserved motif, and the stringency of the hybridization or amplification will determine whether the probe identifies only naturally occurring sequences encoding MTRP, allelic variants, or related sequences.
Probes may also be used for the detection of related sequences, and may have at least 50%
sequence identity to any of the MTRP encoding sequences. The hybridization probes of the subject invention may be DNA or RNA and may be derived from the sequence of SEQ ID
N0:18-34 or from genomic sequences including promoters, enhancers, and introns of the MTRP
gene.
Means for producing specific hybridization probes for DNAs encoding MTRP
include the cloning of polynucleotide sequences encoding MTRP or MTRP derivatives into vectors for the production of mRNA probes. Such vectors are known in the art, are commercially available, and may be used to synthesize RNA probes in vitro by means of the addition of the appropriate RNA
polymerases and the appropriate labeled nucleotides. Hybridization probes maybe labeled by a variety of reporter groups, for example, by radionuclides such as''-P or'sS, or by enzymatic labels, such as alkaline phosphatase coupled to the probe via avidin/biotin coupling systems, and the like.
Polynucleotide sequences encoding MTRP may be used for the diagnosis of disorders associated with expression of MTRP. Examples of such disorders include, but are not limited to, a membrane transport disorder such as cystinuria, dibasicaminoaciduria, hypercystinuria, lysinuria, hartnup disease, tryptophan malabsorption, methionine malabsorption, histidinuria, iminoglycinuria, dicarboxylicaminoaciduria, cystinosis, renal glycosuria, glucose-galactose malabsorption, familial hypercholesterolemia, hypouricemia, familial hypophophatemic rickets, congenital chloridorrhea, cystic fibrosis, familial goiter, distal renal tubular acidosis, Menkes' disease, lethal diarrhea, nephrogenic diabetes insipidus, juvenile pernicious anemia, folate malabsorption, adrenoleukodystrophy, hereditary myoglobinuria, Zellweger syndrome, hyperinsulinemic hypoglycemia, akinesia, amyotrophic lateral sclerosis, ataxia telangiectasia, cystic fibrosis, Becker's IS muscular dystrophy, Bell's palsy, Charcot-Marie Tooth disease, diabetes mellitus, diabetes insipidus, diabetic neuropathy, Duchenne muscular dystrophy, hyperkalemic periodic paralysis, normokalemic periodic paralysis, Parkinson's disease, malignant hyperthermia, multidrug resistance, myasthenia gravis, myotonic dystrophy, catatonia, tardive dyskinesia, dystonias, peripheral neuropathy, cerebral neoplasms, and prostate cancer; a cardiac disorder associated with transport such as angina, bradyarrythmia, tachyarrythmia, hypertension, Long QT syndrome, myocarditis, cardiomyopathy, nemaline myopathy, centronuclear myopathy, lipid myopathy, mitochondria) myopathy, thyrotoxic myopathy, ethanol myopathy, dermatomyositis, inclusion body myositis, infectious myositis, and polymyositis; a neurological disorder associated with transport such as Alzheimer's disease, amnesia, bipolar disorder, dementia, depression, epilepsy, Tourette's disorder, paranoid psychoses, and schizophrenia; and an other disorder associated with transport such as neurofibromatosis, postherpetic neuralgia, trigeminal neuropathy, sarcoidosis, sickle cell anemia, Wilson's disease, cataracts, infertility, pulmonary artery stenosis, sensorineural autosomal deafness, hyperglycemia, hypoglycemia, Grave's disease, goiter, Cushing's disease, Addison's disease, glucose-galactose malabsorption syndrome, and hypercholesterolemia; an immune/inflammatory disorder such as acquired immunodeficiency syndrome (AIDS), Addison's disease, adult respiratory distress syndrome, allergies, ankylosing spondylitis, amyloidosis, anemia, asthma, atherosclerosis, autoimmune hemolytic anemia, autoimrriune thyroiditis, autoimmune polyenodocrinopathy-candidiasis-ectodermal dystrophy (APECED), bronchitis, cholecystitis, contact dermatitis, Crohn's disease, atopic dermatitis, dermatomyositis, diabetes mellitus, emphysema, episodic lymphopenia with lymphocytotoxins; erythroblastosis fetalis, erythema nodosum, atrophic gastritis, glomerulonephritis, Goodpasture's syndrome, gout, Graves' disease, Hashimoto s thyroiditis, hypereosinophilia, irritable bowel syndrome, multiple sclerosis, myasthenia gravis, myocardial or pericardial inflammation, osteoarthritis, osteoporosis, pancreatitis, polymyositis, psoriasis, Reiter's syndrome, rheumatoid arthritis, scleroderma, Sjogren's syndrome, systemic anaphylaxis, systemic lupus erythematosus, systemic sclerosis, thrombocytopenic purpura, ulcerative colitis, uveitis, Werner syndrome, complications of cancer, hemodialysis, and extracorporeal circulation; a viral, bacterial, fungal, parasitic, protozoal, or helminthic infection; and trauma; a cell proliferative disorder such as actinic keratosis, arteriosclerosis, atherosclerosis, bursitis, cirrhosis, hepatitis, mixed connective tissue disease (MCTD), myelofibrosis, paroxysmal nocturnal hemoglobinuria, polycythemia vera, psoriasis, primary thrombocythemia; and a cancer including adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, teratocarcinoma; and, in particular, a cancer of the adrenal gland, bladder, bone, bone marrow, brain, breast, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, penis, prostate, salivary glands, skin, spleen, testis, thymus, thyroid, and uterus. The polynucleotide sequences encoding MTRP may be used in Southern or northern analysis, dot blot, or other membrane-based technologies; in PCR
technologies; in dipstick, pin, and muttiformat ELISA-like assays; and in microarrays utilizing fluids or tissues from patients to detect altered MTRP expression. Such qualitative or quantitative methods are well known in the art.
In a particular aspect, the nucleotide sequences encoding MTRP may be useful in assays that detect the presence of associated disorders, particularly those mentioned above. The nucleotide sequences encoding MTRP may be labeled by standard methods and added to a fluid or tissue sample from a patient under conditions suitable for the formation of hybridization complexes. After a suitable incubation period, the sample is washed and the signal is quantified and compared with a standard value. If the amount of signal in the patient sample is significantly altered in comparison to a control sample then the presence of altered levels of nucleotide sequences encoding MTRP in the sample indicates the presence of the associated disorder. Such assays may also be used to evaluate the efficacy of a particular therapeutic treatment regimen in animal studies, in clinical trials, or to monitor the treatment of an individual patient.
In order to provide a basis for the diagnosis of a disorder associated with expression of MTRP, a normal or standard profile for expression is established. This may be accomplished by combining body fluids or cell extracts taken from normal subjects, either animal or human, with a sequence, or a fragment thereof, encoding MTRP, under conditions suitable for hybridization or amplification. Standard hybridization may be quantified by comparing the values obtained from normal subjects with values from an experiment in which a known amount of a substantially purified polynucleotide is used. Standard values obtained in this manner may be compared with values obtained from samples from patients who are symptomatic for a disorder.
Deviation from standard values is used to establish the presence of a disorder.
Once the presence of a disorder is established and a treatment protocol is initiated, hybridization assays may be repeated on a regular basis to determine if the level of expression in the patient begins to approximate that which is observed in the normal subject.
The results obtained from successive assays may be used to show the efficacy of treatment over a period ranging from several days to months.
p With respect to cancer, the presence of an abnormal amount of transcript (either under- or overexpressed) in biopsied tissue from an individual may indicate a predisposition for the development of the disease, or may provide a means for detecting the disease prior to the appearance of actual clinical symptoms. A more definitive diagnosis of this type may allow health professionals to employ preventative measures or aggressive treatment earlier thereby preventing the development I S or further progression of the cancer.
Additional diagnostic uses for oligonucleotides designed from the sequences encoding MTRP
may involve the use of PCR. These oligomers may be chemically synthesized, generated enzymatically, or produced in vitro. Oligomers will preferably contain a fragment of a polynucleotide encoding MTRP, or a fragment of a polynucleotide complementary to the polynucleotide encoding 20 MTRP, and will be employed under optimized conditions for identification of a specific gene or condition. Oligomers may also be employed under less stringent conditions for detection or quantification of closely related DNA or RNA sequences.
Methods which may also be used to quantify the expression of MTRP include radiolabeling or biotinylating nucleotides, coamplification of a control nucleic acid, and interpolating results from 25 standard curves. (See, e.g., Melby, P.C. et al. ( 1993) J. Immunol. Methods 159:235-244; Duplaa, C.
et al. (1993) Anal. Biochem. 212:229-236.) The speed of quantitation of multiple samples may be accelerated by running the assay in a high-throughput format where the oligomer of interest is presented in various dilutions and a spectrophotometric or colorimetric response gives rapid quantitation.
30 In further embodiments, oligonucleotides or longer fragments derived from any of the polynucleotide sequences described herein may be used as targets in a microarray. The microarray can be used to monitor the expression level of large numbers of genes simultaneously and to identify genetic variants, mutations, and polymorphisms. This information may be used to determine gene function, to understand the genetic basis of a disorder, to diagnose a disorder, and to develop and monitor the activities of therapeutic agents.
Microarrays may be prepared, used, and analyzed using methods known in the art. (See, e.g., Bfennan, T.M. et at. (1995) U.S. Patent No. 5,474,796; Schena, M. et al.
(1996) Proc. Natl. Acad. Sci.
USA 93:10614-10619; Baldeschweiler et al. (1995) PCT application W095/251116;
Shalon, D. et al.
(1995) PCT application W095/35505; Heller, R.A. et al. (1997) Proc. Natl.
Acad. Sci. USA 94:2150-2155; and Heller, M.J. et al. ( 1997) U.S. Patent No. 5,605,662.) In another embodiment of the invention, nucleic acid sequences encoding MTRP
may be used to generate hybridization probes useful in mapping the naturally occurring genomic sequence. The sequences may be mapped to a particular chromosome, to a specific region of a chromosome, or to artificial chromosome constructions, e.g., human artificial chromosomes (HACs), yeast artificial chromosomes (PACs), bacterial artificial chromosomes (BACs), bacterial P1 constructions, or single chromosome cDNA libraries. (See, e.g., Harrington, J.J. et al. (1997) Nat.
Genet. 15:345-355; Price, C.M. (1993) Blood Rev. 7:127-134; and Trask, B.J. (1991) Trends Genet. 7:149-154.) Fluorescent in situ hybridization (FISH) may be correlated with other physical chromosome t5 mapping techniques and genetic map data. (See, e.g., Heinz-Ulrich, et al.
(1995) in Meyers, supra, pp. 965-968.) Examples of genetic map data can be found in various scientific journals or at the Online Mendelian Inheritance in Man (OMIM) World Wide Web site. Correlation between the location of the gene encoding MTRP on a physical chromosomal map and a specific disorder, or a predisposition to a specific disorder, may help define the region of DNA
associated with that disorder.
The nucleotide sequences of the invention may be used to detect differences in gene sequences among normal, carrier, and affected individuals.
In situ hybridization of chromosomal preparations and physical mapping techniques, such as linkage analysis using established chromosomal markers, may be used for extending genetic maps.
Often the placement of a gene on the chromosome of another mammalian species, such as mouse, may reveal associated markers even if the number or arm of a particular human chromosome is not known. New sequences can be assigned to chromosomal arms by physical mapping.
This provides valuable information to investigators searching for disease genes using positional cloning or other gene discovery techniques. Once the disease or syndrome has been crudely localized by genetic linkage to a particular genomic region, e.g., ataxia-telangiectasia to 11 q22-23, any sequences mapping to that area may represent associated or regulatory genes for further investigation. (See, e.g., Gatti, R.A. et al. ( 1988) Nature 336:577-580.) The nucleotide sequence of the subject invention may also be used to detect differences in the chromosomal location due to translocation, inversion, etc., among normal, carrier, or affected individuals.
In another embodiment of the invention, MTRP, its catalytic or immunogenic fragments, or oligopeptides thereof can be used for screening libraries of compounds in any of a variety of drug screening techniques. The fragment employed in such screening may be free in solution, affixed to a solid support, borne on a cell surface, or located intracellularly. The formation of binding complexes between MTRP and the agent being tested may be measured.
Another technique for drug screening provides for high throughput screening of compounds having suitable binding affinity to the protein of interest. (See, e.g., Geysen, et al. ( 1984) PCT
application W084/03564.) Ln this method, large numbers of different small test compounds are synthesized on a solid substrate. The test compounds are reacted with MTRP, or fragments thereof, and washed. Bound MTRP is then detected by methods well known in the art.
Purified MTRP can also be coated directly onto plates for use in the aforementioned drug screening techniques.
Alternatively, non-neutralizing antibodies can be used to capture the peptide and immobilize it on a solid support.
In another embodiment, one may use competitive drug screening assays in which neutralizing antibodies capable of binding MTRP specifically compete with a test compound for binding MTRP.
In this manner, antibodies can be used to detect the presence of any peptide which shares one or more antigenic determinants with MTRP.
In additional embodiments, the nucleotide sequences which encode MTRP may be used in any molecular biology techniques that have yet to be developed, provided the new techniques rely on properties of nucleotide sequences that are currently known, including, but not limited to, such properties as the triplet genetic code and specific base pair interactions.
Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. The following preferred specific embodiments are, therefore, to be construed as merely illustrative, and not limitative ofthe remainder of the disclosure in any way whatsoever.
The disclosures of all patents, applications, and publications mentioned above and below, in particular U.S. Ser. No. [Attorney Docket No. PF-0633 P, filed November 4, 1998], U.S. Ser. No.
[Attorney Docket No. PF-0645 P, filed November 24, 1998], U.S. Ser. No.
[Attorney Docket No. PF-0657 P, filed December 22, 1998], and U.S. Ser. No. 60/121,896, are hereby expressly incorporated by reference.
EXAMPLES
I. Construction of cDNA Libraries RNA was purchased from Clontech or isolated from tissues described in Table 4.
Some tissues were homogenized and lysed in guanidinium isothiocyanate, while others were homogenized and lysed in phenol or in a suitable mixture of denaturants, such as TRIZOL
(Life Technologies), a monophasic solution of phenol and guanidine isothiocyanate. The resulting lysates were centrifuged over CsCI cushions or extracted with chloroform. RNA was precipitated from the lysates with either isopropanoi or sodium acetate and ethanol, or by other routine methods.
Phenol extraction and precipitation of RNA were repeated as necessary to increase RNA
purity. In some cases, RNA was treated with DNase. For most libraries, poly(A+) RNA was isolated using oligo d(T)-coupled paramagnetic particles (Promega), OLIGOTEX latex particles (QIAGEN, Chatsworth CA), or an OLIGOTEX mRNA purification kit (QIAGEN). Alternatively, RNA was isolated directly from tissue lysates using other RNA isolation kits, e.g., the POLY(A)PURE mRNA
purification kit (Ambion, Austin TX).
In some cases, Stratagene was provided with RNA and constructed the corresponding cDNA
libraries. Otherwise, cDNA was synthesized and cDNA libraries were constructed with the UNIZAP
vector system (Stratagene) or SUPERSCRIPT plasmid system (Life Technologies), using the recommended procedures or similar methods known in the art. (See, e.g., Ausubel, 1997, supra, units 5.1-6.6.) Reverse transcription was initiated using oligo d(T) or random primers. Synthetic oligonueleotide adapters were ligated to double stranded cDNA, and the cDNA
was digested with the appropriate restriction enzyme or enzymes. For most libraries, the cDNA was size-selected (300-1000 bp) using SEPHACRYL 51000, SEPHAROSE CL2B, or SEPHAROSE CL4B column chromatography (Amersham Pharmacia Biotech) or preparative agarose gel electrophoresis. cDNAs were ligated into compatible restriction enzyme sites of the polylinker of a suitable plasmid, e.g., PBLUESCRIPT plasmid (Stratagene), PSPORT1 plasmid (Life Technologies), or pINCY (Incyte Pharmaceuticals, Palo Alto CA). Recombinant plasmids were transformed into competent E. coli cells including XLI-Blue, XLI-BIueMRF, or SOLR from Stratagene or DHSa, DHIOB, or ElectroMAX DHIOB from Life Technologies.
II. Isolation of cDNA Clones Plasmids were recovered from host cells by in vivo excision using the UNIZAP
vector system (Stratagene) or by cell lysis. Plasmids were purified using at least one of the following: a Magic or WIZARD Minipreps DNA purification system (Promega); an AGTC Miniprep purification kit (Edge Biosystems, Gaithersburg MD); and QIAWELL 8 Plasmid, QIAWELL 8 Plus Plasmid, Ultra Plasmid purification systems or the R.E.A.L. PREP 96 plasmid purification kit from QIAGEN.
Following precipitation, plasmids were resuspended in 0.1 ml of distilled water and stored, with or without lyophiiization, at 4°C.
Alternatively, plasmid DNA was amplified from host cell lysates using direct link PCR in a high-throughput format(Rao, V.B. (1994) Anal. Biochem. 216:1-14): Host cell lysis and thermal cycling steps were carried out in a single reaction mixture. Samples were proces"sed and stored in 384-well plates, and the concentration of amplified plasmid DNA was quantified fluorometrically using PICOGREEN dye (Molecular Probes, Eugene OR) and a FLUOROSKAN II
fluorescence scanner (Labsystems Oy, Helsinki, Finland).
III. Sequencing and Analysis cDNA sequencing reactions were processed using standard methods or high-throughput instrumentation such as the ABI CATALYST 800 (Perkin-Elmer) thermal cycler or the PTC-200 thermal cycler (MJ Research) in conjunction with the HYDRA microdispenser (Robbins Scientific) or the MICROLAB 2200 (Hamilton) liquid transfer system. cDNA sequencing reactions were prepared using reagents provided by Amersham Pharmacia Biotech or supplied in ABl sequencing kits such as the ABI PR1SM BIGDYE Terminator cycle sequencing ready reaction kit (Perkin-Elmer).
Electrophoretic separation of cDNA sequencing reactions and detection of labeled polynucleotides were carried out using the MEGABACE 1000 DNA sequencing system (Molecular Dynamics}; the IS ABI PRISM 373 or 377 sequencing system (Perkin-Elmer) in conjunction with standard ABI
protocols and base calling software; or other sequence analysis systems known in the art. Reading frames within the cDNA sequences were identified using standard methods (reviewed in Ausubel, 1997, supra, unit 7.7). Some of the cDNA sequences were selected for extension using the techniques disclosed in Example V.
The polynucleotide sequences derived from cDNA sequencing were assembled and analyzed using a combination of software programs which utilize algorithms well known to those skilted in the art. Table S summarizes the tools, programs, and algorithms used and provides applicable descriptions, references, and threshold parameters. The first column of Table 5 shows the tools, programs, and algorithms used, the second column provides brief descriptions thereof, the third column presents appropriate references, all of which are incorporated by reference herein in their entirety, and the fourth column presents, where applicable, the scores, probability values, and other parameters used to evaluate the strength of a match between two sequences (the higher the score, the greater the homology between two sequences). Sequences were analyzed using MACDNASIS PRO
software (Hitachi Software Engineering, South San Francisco CA) and LASERGENE
software (DNASTAR). Polynucleotide and polypeptide sequence alignments were generated using the default parameters specified by the clustal algorithm as incorporated into the MEGALIGN multisequence alignment program (DNASTAR), which also calculates the percent identity between aligned sequences.
The polynucleotide sequences were validated by removing vector, linker, and polyA

sequences and by masking ambiguous bases, using algorithms and programs based on BLAST, dynamic programing, and dinucleotide nearest neighbor analysis. The sequences were then queried against a selection of public databases such as the GenBank primate, rodent, mammalian, vertebrate, and eukaryote databases, and BLOCKS, PRINTS, DOMO, PRODOM, and PFAM to acquire annotation using programs based on BLAST, FASTA, and BLIMPS. The sequences were assembled into full length polynucleotide sequences using programs based on Phred, Phrap, and Consed, and were screened for open reading frames using programs based on GeneMark, BLAST, and FASTA.
The full length polynucleotide sequences were translated to derive the corresponding full length amino acid sequences, and these full length sequences were subsequently analyzed by querying against databases such as the GenBank databases (described above), SwissProt, BLOCKS, PRINTS, DOMO, PRODOM, Prosite, and Hidden Markov Model (HMM~based protein family databases such as PFAM. HMM is a probabilistic approach which analyzes consensus primary structures of gene families. (See, e.g., Eddy, S.R. (1996) Curr. Opin. Struct. Biol. 6:361-365.) The programs described above for the assembly and analysis of full length polynucleotide and amino acid sequences were also used to identify polynucleotide sequence fragments from SEQ ID
N0:18-34. Fragments from about 20 to about 4000 nucleotides which are useful in hybridization and amplification technologies were described in The Invention section above.
IV. Northern Analysis Northern analysis is a laboratory technique used to detect the presence of a transcript of a gene and involves the hybridization of a labeled nucleotide sequence to a membrane on which RNAs from a particular cell type or tissue have been bound. (See, e.g., Sambrook, s. upra, ch. 7; Ausubel, 1995, suara, ch. 4 and '16.) Analogous computer techniques applying BLAST were used to search for identical or related molecules in nucleotide databases such as GenBank or LIFESEQ (lncyte Pharmaceuticals). This analysis is much faster than multiple membrane-based hybridizations. In addition, the sensitivity of the computer search can be modified to determine whether any particular match is categorized as exact or similar. The basis ofthe search is the product score, which is defined as:
sequence identity x % maximum BLAST score The product score takes into account both the degree of similarity between two sequences and the length of the sequence match. For example, with a product score of 40, the match will be exact within a 1% to 2% error, and, with a product score of 70, the match will be exact.
Similar molecules are usually identified by selecting those which show product scores between i 5 and 40, although lower scores may identify related molecules.

The results of northern analyses are reported as a percentage distribution of libraries in which the transcript encoding MTRP occurred. Analysis involved the categorization oi=cDNA libraries by organ/tissue and disease. The organ/tissue categories included cardiovascular, dermatologic, developmental, endocrine, gastrointestinal, hematopoietic/immune, musculoskeletal, nervous, reproductive, and urologic. The disease/condition categories included cancer, inflammation, trauma, cell proliferation, neurological, and pooled. For each category, the number of libraries expressing the sequence of interest was counted and divided by the total number of libraries across all categories.
Percentage values of tissue-specific and disease- or condition-specific expression are reported in Table 3 V. Extension of MTRP Encoding Polynucleotides The full length nucleic acid sequences of SEQ ID NO: I 8-34 were produced by extension of an appropriate fragment of the full length molecule using oligonucleotide primers designed from this fragment. One primer was synthesized to initiate 5' extension of the known fragment, and the other primer, to initiate 3' extension of the known fragment. The initial primers were designed using OLIGO 4.06 software (National Biosciences}, or another appropriate program, to be about 22 to 30 nucleotides in length, to have a GC content of about 50% or more, and to anneal to the target sequence at temperatures of about 68°C to about 72°C. Any stretch of nucleotides which would result in hairpin structures and primer-primer dimerizations was avoided.
Selected human cDNA libraries were used to extend the sequence. If more than one extension was necessary or desired, additional or nested sets of primers were designed.
High fidelity amplification was obtained by PCR using methods well known in the art. PCR
was performed in 96-well plates using the PTC-200 thermal cycler (MJ Research, Inc.). The reaction mix contained DNA template, 200 nmol of each primer, reaction buffer containing Mg=', (NH,,)ZS04, and ~3-mercaptoethanol, Taq DNA polymerase (Amersham Pharmacia Biotech), ELONGASE enzyme (Life Technologies), and Pfu DNA polymerase (Stratagene), with the following parameters for primer pair PCI A and PCI B: Step 1: 94°C, 3 min; Step 2: 94°C, 15 sec;
Step 3: 60°C, 1 min; Step 4: 68°C, 2 min; Step 5: Steps 2, 3, and 4 repeated 20 times; Step 6: 68°C, 5 min; Step 7: storage at 4°C. In the alternative, the parameters for primer pair T7 and SK+ were as follows: Step 1: 94°C, 3 min; Step 2:
94°C, 15 sec; Step 3: 57°C, 1 min; Step 4: 68°C, 2 min;
Step 5: Steps 2, 3, and 4 repeated 20 times;
Step 6: 68°C, 5 min; Step 7: storage at 4°C.
The concentration of DNA in each well was determined by dispensing 100 pl PICOGREEN
quantitation reagent (0.25% (v/v) PICOGREEN; Molecular Probes, Eugene OR) dissolved in 1 X TE
and 0.5 pl of undiluted PCR product into each well of an opaque fluorimeter plate (Corning Costar, Acton MA), allowing the DNA to hind to the reagent. The plate was scanned in a Fluoroskan II

(Labsystems Oy, Helsinki, Finland) to measure the fluorescence of the sample and to quantify the concentration of DNA. A 5 ~cl to 10 ~cl aliquot of the reaction mixture was analyzed by electrophoresis on a 1 % agarose mini-gel to determine which reactions were successful in extending the sequence.
The extended nucleotides were desalted and concentrated, transferred to 384-well plates, digested with CviJI cholera virus endonuclease (Molecular Biology Research, Madison WI), and sonicated or sheared prior to relegation into pUC 18 vector (Amersham Pharmacia Biotech). For shotgun sequencing, the digested nucleotides were separated on low concentration (0.6 to 0.8%) agarose gels, fragments were excised, and agar digested with Agar ACE
(Promega). Extended clones were relegated using T4 ligase (New England Biolabs, Beverly MA) into pUC I 8 vector (Amersham Pharmacia Biotech), treated with Pfu DNA polymerase (Stratagene) to fill-in restriction site overhangs, and transfected into competent E. coli cells. Transformed cells were selected on antibiotic-containing media, individual colonies were picked and cultured overnight at 37°C in 384-well plates in LB/2x carb liquid media.
The cells were lysed, and DNA was amplified by PCR using Taq DNA polymerase (Amersham Pharmacia Biotech) and Pfu DNA polymerase (Stratagene) with the following parameters: Step 1: 94°C, 3 min; Step 2: 94°C, IS sec; Step 3:
60°C, 1 min; Step 4: 72°C, 2 min;
Step 5: steps 2, 3, and 4 repeated 29 times; Step 6: 72°C, 5 min; Step 7: storage at 4°C. DNA was quantified by PICOGREEN reagent (Molecular Probes) as described above. Samples with low DNA
recoveries were reamplified using the same conditions as described above.
Samples were diluted with 20% dimethysulfoxide (1:2, v/v), and sequenced using DYENAMIC energy transfer sequencing primers and the DYENAMIC DIRECT kit (Arnersham Pharmacia Biotech) or the ABI
PRISM
BIGDYE Terminator cycle sequencing ready reaction kit (Perkin-Elmer).
In like manner, the nucleotide sequences of SEQ ID NO:I 8-34 are used to obtain S' regulatory sequences using the procedure above, oligonucleotides designed for such extension, and an appropriate genomic library.
VI. Labeling and Use of Individual Hybridization Probes Hybridization probes derived from SEQ ID N0:18-34 are employed to screen cDNAs, genomic DNAs, or mRNAs. Although the labeling of oligonucleotides, consisting of about 20 base pairs, is specifically described, essentially the same procedure is used with larger nucleotide fragments. Oligonucleotides are designed using state-of the-art software such as OLIGO 4.06 software (National Biosciences) and labeled by combining 50 pmol of each oligomer, 250 ~Ci of [y-'ZP] adenosine triphosphate (Amersham Pharmacia Biotech), and T4 polynucleotide kinase (DuPont NEN, Boston MA). The labeled oligonucleotides are substantially purified using a SEPHADEX G-25 superfine size exclusion dextran bead column (Amersham Pharmacia Biotech).
An aliquot containing 10' counts per minute of the labeled probe is used in a typic"al membrane-based hybridization analysis of human genomic DNA digested with one of the following endonucleases:
Ase I, Bgl II, Eco RI, Pst I, Xba I, or Pvu II (DuPont NEN).
The DNA from each digest is fractionated on a 0.7% agarose gel and transferred to nylon membranes (Nytran Plus, Schleicher & Schuell, Durham NH). Hybridization is carried out for 16 hours at 40°C. To remove nonspecific signals, blots are sequentially washed at room temperature under conditions of up to, for example, 0.1 x saline sodium citrate and 0.5%
sodium dodecyl sulfate.
Hybridization patterns are visualized using autoradiography or an alternative imaging means and compared.
VII. Microarrays A chemical coupling procedure and an ink jet device can be used to synthesize array elements -)? -on the surface of a substrate. (See, e.g., Baldeschweiler, supra.) An array analogous to a dot or slot blot may also be used to arrange and link elements to the surface of a substrate using thermal, UV, chemical, or mechanical bonding procedures. A typical array may be produced by hand or using available methods and machines and contain any appropriate number of elements.
After hybridization, nonhybridized probes are removed and a scanner used to determine the levels and patterns of fluorescence. The degree of complementarity and the relative abundance of each probe which hybridizes to an element on the microarray may be assessed through analysis of the scanned images.
Full-length cDNAs, Expressed Sequence Tags (ESTs), or fragments thereof may comprise the elements of the microarray. Fragments suitable for hybridization can be selected using software well known in the art such as LASERGENE software (DNASTAR). Full-length cDNAs, ESTs, or fragments thereof corresponding to one of the nucleotide sequences of the present invention, or selected at random from a cDNA library relevant to the present invention, are arranged on an appropriate substrate, e.g., a glass slide. The cDNA is fixed to the slide using, e.g., UV cross-linking followed by thermal and chemical treatments and subsequent drying. {See, e.g., Schena, M. et al.
(1995) Science 270:467-470; Shalon, D. et al. (1996) Genome Res. 6:639-645.) Fluorescent probes are prepared and used for hybridization to the elements on the substrate. The substrate is analyzed by procedures described above.
VIII. Complementary Polynucleotides Sequences complementary to the MTRP-encoding sequences, or any parts thereof, are used to detect, decrease, or inhibit expression of naturally occurring MTRP.
Although use of oligonucleotides comprising from about 15 to 30 base pairs is described, essentially the same procedure is used with smaller or with larger sequence fragments. Appropriate oligonucleotides are designed using OLIGO 4.06 software (National Biosciences) and the coding sequence of MTRP. To inhibit transcription, a complementary oligonucleotide is designed from the most unique 5' sequence and used to prevent promoter binding to the coding sequence. To inhibit translation, a complementary oligonucleotide is designed to prevent ribosomal binding to the MTRP-encoding transcript.
IX. Expression of MTRP
Expression and purification of MTRP is achieved using bacterial or virus-based expression systems. For expression of MTRP in bacteria, cDNA is subcloned into an appropriate vector containing an antibiotic resistance gene and an inducible promoter that directs high levels of cDNA
transcription. Examples of such promoters include, but are not limited to, the trp-lac (tac) hybrid promoter and the TS or T7 bacteriophage promoter in conjunction with the lac operator regulatory element. Recombinant vectors are transformed into suitable bacterial hosts, e.g., BL21(DE3).
Antibiotic resistant bacteria express MTRP upon induction with isopropyl beta-D-thiogalactopyranoside (IPTG). Expression of MTRP in eukaryotic cells is achieved by infecting insect or mammalian cell lines with recombinant Autographica californica nuclear polyhedrosis virus (AcMNPV), commonly known as bacuiovirus. The nonessential polyhedrin gene of baculovirus is replaced with cDNA encoding MTRP by either homologous recombination or bacterial-mediated transposition involving transfer plasmid intermediates. Viral infectivity is maintained and the strong polyhedrin promoter drives high levels of cDNA transcription. Recombinant baculovirus is used to infect Snodoptera frugi~erda (Sf9) insect cells in most cases, or human hepatocytes, in some cases.
Infection of the latter requires additional genetic modifications to baculovirus. (See Engelhard, E.K.
et al. (1994) Proc. Natl. Acad. Sci. USA 91:3224-3227; Sandig, V. et al.
(1996) Hum. Gene Ther.
7:1937-1945.) In most expression systems, MTRP is synthesized as a fusion protein with, e.g., glutathione S-transferase (GST) or a peptide epitope tag, such as FLAG or 6-His, permitting rapid, single-step, affinity-based purification of recombinant fusion protein from crude cell lysates. GST, a 26-kilodalton enzyme from Schistosoma ja onp icum, enables the purification of fusion proteins on immobilized glutathione under conditions that maintain protein activity and antigenicity (Amersham Pharmacia Biotech). Following purifECation, the GST moiety can be proteolytically cleaved from MTRP at specii~ically engineered sites. FLAG, an 8-amino acid peptide, enables immunoaffinity purification using commercially available monoclonal and polyclonal anti-FLAG
antibodies (Eastman Kodak). 6-His, a stretch of six consecutive histidine residues, enables purification on metal-chelate resins (QIAGEN). Methods for protein expression and purification are discussed in Ausubel ( 1995, supra, ch. 10 and 16). Purified MTRP obtained by these methods can be used directly in the following activity assay, X. Demonstration of MTRP Activity ATPase activity associated with MTRP can be measured by hydrolysis of radiolabeled ATP-[y-''-P], separation of the hydrolysis products by chromatographic methods, and quantitation of the recovered''-P using a scintillation counter. The reaction mixture contains ATP-[y-3'P] and varying amounts of MTRP in a suitable buffer incubated at 37°C for a suitable period of time. The reaction is terminated by acid precipitation with trichloroacetic acid and then neutralized with base, and an aliquot of the reaction mixture is subjected to membrane or filter paper-based chromatography to separate the reaction products. The amount of'ZP liberated is counted in a scintillation counter. The amount of radioactivity recovered is proportional to the ATPase activity of MTRP in the assay.
MTRP transport activity is assayed by measuring uptake of labeled substrates into Xenopus laevis oocytes. Oocytes at stages V and VI are injected with MTRP mRNA (10 ng per oocyte) and incubated for 3 days at 18°C in OR2 medium (82.SmM NaCI, 2.5 mM KCI, 1mM CaCI,, 1mM
MgCI,, 1 mM Na~HP04, 5 mM Hepes, 3.8 mM NaOH , SOp.g/ml gentamycin, pH 7.8) to allow expression of MTRP protein. Oocytes are then transferred to standard uptake medium ( 100mM NaCI, 2 mM KCI, 1mM CaCl2, 1mM MgClz, 10 mM Hepes/Tris, pH 7.5). Uptake of various substrates (e.g., amino acids, sugars, drugs, and neurotransmitters) is initiated by adding a'H-labeled substrate to the oocytes. After 30 minutes of incubation, uptake is terminated by washing the oocytes three times in Na+-free medium. Incorporation of 'H is measured, and compared with controls. MTRP
transport activity is proportional to the level of internalized'H-labeled substrate.
XI. Functional Assays MTRP function is assessed by expressing the sequences encoding MTRP at physiologically elevated levels in mammalian cell culture systems. cDNA is subcloned into a mammalian expression vector containing a strong promoter that drives high levels of cDNA
expression. Vectors of choice include pCMV SPORT (Life Technologies) and pCR3.1 (Invitrogen, Carlsbad CA), both of which contain the cytomegalovirus promoter. 5-10 ,ug of recombinant vector are transiently transfected into a human cell line, for example, an endothelial or hematopoietic cell line, using either liposome formulations or electroporation. 1-2 ug of an additional plasmid containing sequences encoding a marker protein are co-transfected. Expression of a marker protein provides a means to distinguish transfected cells from nontransfected cells and is a reliable predictor of cDNA expression from the recombinant vector. Marker proteins of choice include, e.g., Green Fluorescent Protein (GFP;
Clontech), CD64, or a CD64-GFP fusion protein. Flow cytometry (FCM), an automated, laser optics-based technique, is used to identify transfected cells expressing GFP or CD64-GFP and to evaluate the apoptotic state of the cells and other cellular properties. FCM detects and quantifies the uptake of fluorescent molecules that diagnose events preceding or coincident with cell death. These events include changes in nuclear DNA content as measured by staining of DNA with piopidium iodide;
changes in cell size and granularity as measured by forward light scatter and 90 degree side light scatter; down-regulation of DNA synthesis as measured by decrease in bromodeoxyuridine uptake;
alterations in expression of cell surface and intracellular proteins as measured by reactivity with specific antibodies; and alterations in plasma membrane composition as measured by the binding of fluorescein-conjugated Annexin V protein to the cell surface. Methods in flow cytometry are discussed in Ormerod, M.G. ( 1994) Flow C ometrv, Oxford, New York NY.
The influence of MTRP on gene expression can be assessed using highly purified populations of cells transfected with sequences encoding MTRP and either CD64 or CD64-GFP.
CD64 and CD64-GFP are expressed on the surface of transfected cells and bind to conserved regions of human immunoglobulin G (IgG). Transfected cells are efficiently separated from nontransfected cells using magnetic beads coated with either human IgG or antibody against CD64 (DYNAL, Lake Success NY). mRNA can be purified from the cells using methods well known by those of skill in the I S art. Expression of mRNA encoding MTRP and other genes of interest can be analyzed by northern analysis or microarray techniques.
XII. Production of MTRP Specific Antibodies MTRP substantially purified using polyacrylamide gel electrophoresis (PAGE;
see, e.g., Harrington, M.G. ( 1990) Methods Enzymol. 182:488-495), or other purification techniques, is used to immunize rabbits and to produce antibodies using standard protocols.
Alternatively, the MTRP amino acid sequence is analyzed using LASERGENE
software (DNASTAR) to determine regions of high immunogenicity, and a corresponding oligopeptide is synthesized and used to raise antibodies by means known to those of skill in the art. Methods for selection of appropriate epitopes, such as those near the C-terminus or in hydrophilic regions are well described in the art. (See, e.g., Ausubel, 1995, su ra, ch. 11.) Typically, oligopeptides of about 15 residues in length are synthesized using an ABI 431A
peptide synthesizer (Perkin-Elmer) using fmoc-chemistry and coupled to KLH
(Sigma-Aldrich, St.
Louis MO) by reaction with N-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) to increase immunogenicity. (See, e.g., Ausubel, 1995, su ra.) Rabbits are immunized with the oligopeptide-KLH complex in complete Freund's adjuvant. Resulting antisera are tested for antipeptide and anti-MTRP activity by, for example, binding the peptide or MTRP to a substrate, blocking with 1% BSA, reacting with rabbit antisera, washing, and reacting with radio-iodinated goat anti-rabbit IgG.
XIII. Purification of Naturally Occurring MTRP Using Specific Antibodies Naturally occurring or recombinant MTRP is substantially purified by immunoaffinity chromatography using antibodies specific for MTRP. An immunoaffinity column is constructed by covalently coupling anti-MTRP antibody to an activated chromatographic resin, such as CNBr-activated SEPHAROSE (Amersham Pharmacia Biotech). After the coupling, the resin is blocked and washed according to the manufacturer's instructions.
Media containing MTRP are passed over the~immunoaffinity column, and the column is washed under conditions that allow the preferential absorbance of MTRP (e.g., high ionic strength buffers in the presence of detergent). The column is eluted under conditions that disrupt antibody/MTRP binding (e.g., a buffer of pH 2 to pH 3, or a high concentration of a chaotrope, such as urea or thiocyanate ion), and MTRP is collected.
XIV. Identification of Molecules Which Interact with MTRP
MTRP, or biologically active fragments thereof, are labeled with '251 Bolton-Hunter reagent.
(See, e.g., Bolton A.E. and W.M. Hunter (1973) Biochem. J. 133:529-539.) Candidate molecules previously arrayed in the wells of a multi-well plate are incubated with the labeled MTRP, washed, and any wells with labeled MTRP complex are assayed. Data obtained using different concentrations of MTRP are used to calculate values for the number, affinity, and association of MTRP with the candidate molecules.
Various modifications and variations of the described methods and systems of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention.
Although the invention has been described in connection with certain embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments.
Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in molecular biology or related fields are intended to be within the scope of the following claims.

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SEQUENCE LISTING
<110> INCYTE PHARMACEUTICALS, INC.
HILLMAN, Jennifer L.
YUE, Henry TANG, Y. Tom LAL, Preeti CORLEY, Neil C.
GUEGLER, Karl J.
BAUGHN, Mariah R.
AZIMZAI, Yalda LU, Dyung Aina M.
<120> MEMBRANE TRANSPORT PROTEINS
<130> PF-0633 PCT
<140> To Be Assigned <141> Herewith <150> 09/186,778; unassigned; 09/200,277; unassigned; 09/221,405;
unassigned; 60/121,896 <151> 1998-11-04; 1998-11-04; 1998-11-24; 1998-11-24; 1998-12-22;
1998-12-22; 1999-02-26 <160> 42 <170> PERL Program <210> 1 <211> 384 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 961344CD1 <400> 1 Met Leu Thr Gly Asp Lys Leu Glu Thr Ala Thr Cys Ile Ala Lys Ser Ser His Leu Val Ser Arg Thr Gln Asp Ile His Ile Phe Arg Gln Val Thr Ser Arg Gly Glu Ala His Leu Glu Leu Asn Ala Phe Arg Arg Lys His Asp Cys Ala Leu Val Ile Ser Gly Asp Ser Leu Glu Val Cys Leu Lys Tyr Tyr Glu His Glu Phe Val Glu Leu Ala Cys Gln Cys Pro Ala Val Val Cys Cys Arg Cys Ser Pro Thr Gln Lys Ala Arg Ile Val Thr Leu Leu Gln Gln His Thr Gly Arg Arg Thr Cys Ala Ile Gly Asp Gly Gly Asn Asp Val Ser Met Ile Gln Ala Ala Asp Cys Gly Ile Gly Ile Glu Gly Lys Glu Gly Lys Gln Ala Ser Leu Ala Ala Asp Phe Ser Ile Thr Gln Phe Arg His Ile Gly Arg Leu Leu Met Val His Gly Arg Asn Ser Tyr Lys Arg Ser Ala Ala Leu Gly Gln Phe Val Met His Arg Gly Leu IIe Ile Ser Thr Met Gln Ala Val Phe Ser Ser Val Phe Tyr Phe Ala Ser Val Pro Leu Tyr Gln Gly Phe Leu Met Val Gly Tyr Ala Thr Ile Tyr Thr Met Phe Pro Val Phe Ser Leu Val Leu Asp Gln Asp Val Lys Pro Glu Met Ala Met Leu Tyr Pro Glu Leu Tyr Lys Asp Leu Thr Lys Gly Arg Ser Leu Ser Phe Lys Thr Phe Leu Ile Trp Val Leu Ile Ser Ile Tyr Gln Gly Gly Ile Leu Met Tyr Gly Ala Leu Val Leu Phe Glu Ser Glu Phe Val His Val Val Ala Ile Ser Phe Thr Ala Leu Ile Leu Thr Glu Leu Leu Met Val Ala Leu Thr Val Arg Thr Trp His Trp Leu Met Val Val Ala Glu Phe Leu Ser Leu Gly Cys Tyr Val Ser Ser Leu Ala Phe Leu Asn Glu Tyr Phe Gly Ile Gly Arg Val Ser Phe Gly Ala Phe Leu Asp Val Ala Phe Ile Thr Thr Val Thr Phe Leu Trp Lys Val Ser Ala Ile Thr Val Val Ser Cys Leu Pro Leu Tyr Val Leu Lys Tyr Leu Arg Arg Lys Leu Ser Pro Pro Ser Tyr Cys Lys Leu Ala Ser <210> 2 <211> 846 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 3128782CD1 <400> 2 Met Pro Lys Ala Pro Lys Gln Gln Pro Pro Glu Pro Glu Trp Ile Gly Asp Gly Glu Ser Thr Ser Pro Ser Asp Lys Val Val Lys Lys Gly Lys Lys Asp Lys Lys Ile Lys Lys Thr Phe Phe Glu Glu Leu -35 40 . 45 Ala Val Glu Asp Lys Gln Ala Gly Glu Glu Glu Lys Val Leu Ly$"

Glu Lys Glu Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Lys Lys Lys Arg Asp Thr Arg Lys Gly Arg Arg Lys Lys Asp Val Asp Asp Asp Gly Glu Glu Lys Glu Leu Met Glu Arg Leu Lys Lys Leu Ser Val Pro Thr Ser Asp G,lu Glu Asp Glu Val Pro Ala Pro Lys Pro Arg Gly Gly Lys Lys Thr Lys Gly Gly Asn Val Phe Ala Ala Leu Ile Gln Asp Gln Ser Glu Glu Glu Glu Glu Glu Glu Lys His Pro Pro Lys Pro Ala Lys Pro Glu Lys Asn Arg Ile Asn Lys Ala Val Ser Glu Glu Gln Gln Pro Ala Leu Lys Gly Lys Lys Gly Lys Glu Glu Lys Ser Lys Gly Lys Ala Lys Pro Gln Asn Lys Phe Ala Ala Leu Asp Asn Glu Glu Glu Asp Lys Glu Glu Glu Ile Ile Lys Glu Lys Glu Pro Pro Lys Gln Gly Lys Glu Lys Ala Lys Lys Ala Glu Gln Gly Ser Glu Glu Glu Gly Glu Gly Glu Glu Glu Glu Glu Glu Gly Gly Glu Ser Lys Ala Asp Asp Pro Tyr Ala His Leu Ser Lys Lys Glu Lys Lys Lys Leu Lys Lys Gln Met Glu Tyr Glu Arg Gln Val Ala Ser Leu Lys Ala Ala Asn Ala Ala Glu Asn Asp Phe Ser Val Ser Gln Ala Glu Met Ser Ser Arg Gln Ala Met Leu Glu Asn Ala Sex Asp Ile Lys Leu Glu Lys Phe Ser Ile Ser Ala His Gly Lys Glu Leu Phe Val Asn Ala Asp Leu Tyr Ile Val Ala Gly Arg Arg Tyr Gly Leu Val Gly Pro Asn Gly Lys Gly Lys Thr Thr Leu Leu Lys His Ile Ala Asn Arg Ala Leu Ser Ile Pro Pro Asn Ile Asp Val Leu Leu Cys Glu Gln Glu Val Val Ala Asp Glu Thr Pro Ala Val Gln Ala Val Leu Arg Ala Asp Thr Lys Arg Leu Lys Leu Leu Glu Glu Glu Arg Arg Leu Gln Gly Gln Leu Glu Gln Gly Asp Asp Thr Ala Ala Glu Arg Leu Glu Lys Val Tyr Glu Glu Leu Arg Ala Thr Gly Ala Ala Ala Ala Glu Ala Lys Ala Arg Arg Ile Leu Ala Gly Leu Gly Phe Asp Pro Glu Met Gln Asn Arg Pro Thr Gln Lys Phe Ser Gly Gly Trp Arg Met Arg Val Ser Leu Ala Arg 455 460 46y Ala Leu Phe Met Glu Pro Thr Leu Leu Met Leu Asp Glu Pro Thr Asn His Leu Asp Leu Asn Ala Val Ile Trp Leu Asn Asn Tyr Leu Gln Gly Trp Arg Lys Thr Leu Leu Ile Val Ser His Asp Gln Gly Phe Leu Asp Asp Val Cys Thr Asp Ile Ile His Leu Asp Ala Gln Arg Leu His Tyr Tyr Arg Gly Asn Tyr Met Thr Phe Lys Lys Met Tyr Gln Gln Lys Gln Lys Glu Leu Leu Lys Gln Tyr Glu Lys Gln Glu Lys Lys Leu Lys Glu Leu Lys Ala Gly Gly Lys Ser Thr Lys Gln Ala Glu Lys Gln Thr Lys Glu Ala Leu Thr Arg Lys Gln Gln 5?5 580 585 Lys Cys Arg Arg Lys Asn Gln Asp Glu Glu Ser Gln Glu Ala Pro Glu Leu Leu Lys Arg Pro Lys Glu Tyr Thr Val Arg Phe Thr Phe Pro Asp Pro Pro Pro Leu Ser Pro Pro Val Leu Gly Leu His Gly Val Thr Phe Gly Tyr Gln Gly Gln Lys Pro Leu Phe Lys Asn Leu Asp Phe Gly Ile Asp Met Asp Ser Arg Ile Cys Ile Val Gly Pro Asn Gly Val Gly Lys Ser Thr Leu Leu Leu Leu Leu Thr Gly Lys Leu Thr Pro Thr His Gly Glu Met Arg Lys Asn His Arg Leu Lys Ile Gly Phe Phe Asn Gln Gln Tyr Ala Glu Gln Leu Arg Met Glu Glu Thr Pro Thr Glu Tyr Leu Gln Arg Gly Phe Asn Leu Pro Tyr Gln Asp Ala Arg Lys Cys Leu Gly Arg Phe Gly Leu Glu Ser His Ala His Thr Ile Gln Ile Cys Lys Leu Ser Gly Gly Gln Lys Ala Arg Val Val Phe Ala Glu Leu Ala Cys Arg Glu Pro Asp Val Leu Ile Leu Asp Glu Pro Thr Asn Asn Leu Asp Ile Glu Ser Ile Asp Ala Leu Gly Glu Ala Ile Asn Glu Tyr Lys Gly Ala Val Ile Val Val Ser His Asp Ala Arg Leu Ile Thr Glu Thr Asn Cys Gln Leu Trp Val Val Glu Glu Gln Ser Val Ser Gln Ile Asp Gly Asp Phe Glu Asp Tyr Lys Arg Glu Val Leu Glu Ala Leu Gly Glu Val Met Val Ser Arg Pro Arg Glu <210> 3 ' <211> 511 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 1720440CD1 <400> 3 Met Glu Asn Arg Asn Glu Phe Val Gly Leu Trp Leu Gly Met Ala Lys Leu Gly Val Glu Ala Ala Leu Ile Asn Thr Asn Leu Arg Arg Asp Ala Leu Leu His Cys Leu Thr Thr Ser Arg Ala Arg Ala Leu Val Phe Gly Ser Glu Met Ala Ser Ala Ile Cys Glu Va1 His Ala Ser Leu Asp Pro Ser Leu Ser Leu Phe Cys Ser Gly Ser Trp Glu Pro Gly Ala Val Pro Pro Ser Thr Glu His Leu Asp Pro Leu Leu Lys Asp Ala Pro Lys His Leu Pro Ser Cys Pro Asp Lys Gly Phe Thr Asp Lys Leu Phe Tyr Ile Tyr Thr Ser Gly Thr Thr Gly Leu Pro Lys Ala Ala Ile Val Val His Ser Arg Tyr Tyr Arg Met Ala Ala Leu Val Tyr Tyr Gly Phe Arg Met Arg Pro Asn Asp Ile Val Tyr Asp Cys Leu Pro Leu Tyr His Ser Ala Gly Asn Ile Val Gly Ile Gly Gln Cys Leu Leu His Gly Met Thr Val Val Ile Arg Lys Lys Phe Ser Ala Ser Arg Phe Trp Asp Asp Cys Ile Lys Tyr Asn Cys Thr Ile Val Gln Tyr Ile Gly Glu Leu Cys Arg Tyr Leu Leu Asn Gln Pro Pro Arg Glu Ala Glu Asn Gln His Gln Val Arg Met Ala Leu Gly Asn Gly Leu Arg Gln Ser Ile Trp Thr Asn Phe Ser Ser Arg Phe His Ile Pro Gln Val Ala Glu Phe Tyr Gly Ala Thr Glu Cys Asn Cys Ser Leu Gly Asn Phe Asp Ser Gln Val Gly Ala Cys Gly Phe Asn Ser Arg Ile Leu Ser Ser Val Tyr Pro Ile Arg Leu Val Arg Val Asn Glu Asp Thr Met Glu Leu Ile Arg Gly Pro Asp Gly Val Cys Ile Pro Cys Gln Pro Gly Glu Pro Gly Gln Leu Val Gly Arg IIe Ile Gln Lys Asp Pro Leu Arg Arg Phe Asp Gly Tyr Leu Asn Gln Gly Ala Asn Asn Lys Lys Ile Ala Lys Asp Val 335 340 . 345 Phe Lys Lys Gly Asp Gln Ala Tyr Leu Thr Gly Asp Val Leu Val_ Met Asp Glu Leu Gly Tyr Leu Tyr Phe Arg Asp Arg Thr Gly Asp 365 3?0 375 Thr Phe Arg Trp Lys Gly Glu Asn Val Sex Thr Thr Glu Val Glu Gly Thr Leu Ser Arg Leu Leu Asp Met Ala Asp Val Ala Val Tyr Gly Val Glu Val Pro Gly Thr Glu Gly Arg Ala Gly Met Ala Ala Val Ala Ser Pro Thr Gly Asn Cys Asp Leu Glu Arg Phe Ala Gln Val Leu Glu Lys Glu Leu Pro Leu Tyr Ala Arg Pro Ile Phe Leu Arg Leu Leu Pro Glu Leu His Lys Thr Gly Thr Tyr Lys Phe Gln Lys Thr Glu Leu Arg Lys Glu Gly Phe Asp Pro Ala Ile Val Lys Asp Pro Leu Phe Tyr Leu Asp Ala Gln Lys Gly Arg Tyr Val Pro Leu Asp Gln Glu Ala Tyr Sex Arg Ile Gln Ala Gly Glu Glu,Lys Leu <210> 4 <211> 718 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2274290CD1 <400> 4 Met Leu Val His Leu Phe Arg Val Gly Ile Arg Gly Gly Pro Phe Pro Gly Arg Leu Leu Pro Pro Leu Arg Phe Gln Thr Phe Ser Ala Val Arg Tyr Ser Asp Gly Tyr Arg Ser Ser Ser Leu Leu Arg Ala Val Ala His Leu Arg Ser Gln Leu Trp Ala His Leu Pro Arg Ala Pro Leu Ala Pro Arg Trp Ser Pro Ser Ala Trp Cys Trp Val Gly Gly Ala Leu Leu Gly Pro Met Val Leu Ser Lys His Pro His Leu Cys Leu Val Ala Leu Cys Glu Ala Glu Glu Ala Pro Pro Ala Ser g5 100 105 Ser Thr Pro His Val Val Gly Ser Arg Phe Asn Trp Lys Leu Phe Trp Gln Phe Leu His Pro His Leu Leu Val Leu Gly Val Ala Val Val Leu Ala Leu Gly Ala Ala Leu Val Asn Val Gln Ile Pro Leu PCTlUS99/26048 140 145 . 150 Leu Leu Gly Gln Leu Val Glu Val Val Ala Lys Tyr Thr Arg Asg His Val Gly Ser Phe Met Thr Glu Ser Gln Asn Leu Ser Thr His Leu Leu Ile Leu Tyr Gly Val Gln Gly Leu Leu Thr Phe Gly Tyr Leu Val Leu Leu Ser His Val Gly Glu Arg Met Ala Val Asp Met Arg Arg Ala Leu Phe Ser Ser Leu Leu Arg Gln Asp Ile Thr Phe Phe Asp Ala Asn Lys Thr Gly Gln Leu Val Ser Arg Leu Thr Thr Asp Val Gln Glu Phe Lys Ser Ser Phe Lys Leu Val Ile Ser Gln Gly Leu Arg Sex Cys Thr Gln Val Ala Gly Cys Leu Val Ser Leu Ser Met Leu Ser Thr Arg Leu Thr Leu Leu Leu Met Val Ala Thr Pro Ala Leu Met Gly Val Gly Thr Leu Met Gly Ser Gly Leu Arg Lys Leu Ser Arg Gln Cys Gln Glu Gln Ile Ala Arg Ala Met Gly Val Ala Asp Glu Ala Leu Gly Asn Val Arg Thr Val Arg Ala Phe Ala Met Glu Gln Arg Glu Glu Glu Arg Tyr Gly Ala Glu Leu Glu Ala Cys Arg Cys Arg Ala Glu Glu Leu Gly Arg Gly Ile Ala Leu Phe Gln Gly Leu Ser Asn Ile Ala Phe Asn Cys Met Val Leu Gly Thr Leu Phe Ile Gly Gly Ser Leu Val Ala Gly Gln Gln Leu Thr Gly Gly Asp Leu Met Ser Phe Leu Val Ala Ser Gln Thr Val Gln Arg Ser Met Ala Asn Leu Ser Val Leu Phe Gly Gln Val Val Arg Gly Leu Ser Ala Gly Ala Arg Val Phe Glu Tyr Met Ala Leu Asn Pro Cys Ile Pro Leu Ser Gly Gly Cys Cys Val Pro Lys Glu Gln Leu Arg Gly Ser Val Thr Phe Gln Asn Val Cys Phe Ser Tyr Pro Cys Arg Pro Gly Phe Glu Val Leu Lys Asp Phe Thr Leu Thr Leu Pro Pro Gly Lys Ile Val Ala Leu Val Gly Gln Ser Gly Gly Gly Lys Thr Thr Val Ala Ser Leu Leu Glu Arg Phe Tyr Asp Pro Thr Ala Gly Val Val Met Leu Asp Gly Arg Asp Leu Arg Thr Leu Asp Pro Ser Trp Leu Arg Gly Gln Val Val G1y Phe Ile Ser Gln Glu Pro VaI Leu Phe Gly Thr Thr ile Met Glu Asn Ile Arg Phe Gly Lys Leu Glu Ala Ser Asp Glu Glu Val Tyr Thr Ala Ala Arg Glu Ala Asn Ala His Glu Phe Ile Thr Ser Phe Pro Glu Gly Tyr Asn Thr Val Val Gly Glu Arg Gly Thr Thr Leu Ser Gly Gly Gln Lys Gln Arg Leu Ala Ile Ala Arg Ala Leu Ile Lys Gln Pro Thr Val Leu Ile Leu Asp Glu Ala Thr Ser Ala Leu Asp Ala Glu Ser Glu Arg Val Val Gln Glu Ala Leu Asp Arg Ala Ser Ala Gly Arg Thr Val Leu Val Ile Ala His Arg Leu Ser Thr Val Arg Gly Ala His Cys Ile Val Val Met Ala Asp Gly Arg Val Trp Glu Ala Gly Thr His Glu Glu Leu Leu Lys Lys Gly Gly Leu Tyr Ala Glu Leu Ile Arg Arg Gln Ala Leu Asp Ala Pro Arg Thr Ala Ala Pro Pro Pro Lys Lys Pro Glu Gly Pro Arg Ser His Gln His Lys Ser <210> 5 <211> 635 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2740029CD1 <400> 5 Met Ser Val Gly Val Ser Thr Ser Ala Pro Leu Ser Pro Thr Ser Gly Thr Ser Val Gly Met Ser Thr Phe Ser Ile Met Asp Tyr Val Val Phe Val Leu Leu Leu Val Leu Ser Leu Ala Ile Gly Leu Tyr His Ala Cys Arg Gly Trp Gly Arg His Thr Val Gly Glu Leu Leu Met Ala Asp Arg Lys Met Gly Cys Leu Pro Val Ala Leu Ser Leu Leu Ala Thr Phe Gln Ser Ala Val Ala Ile Leu Gly Val Pro Ser Glu Ile Tyr Arg Phe Gly Thr Gln Tyr Trp Phe Leu Gly Cys Cys g5 100 105 Tyr Phe Leu Gly Leu Leu Ile Pro Ala His Ile Phe Ile Pro Val Phe Tyr Arg Leu His Leu Thr Ser Ala Tyr Glu Tyr Leu Glu Leu Arg Phe Asn Lys Thr Val Arg Val Cys Gly Thr Val Thr Phe Ile Phe Gln Met Val Ile Tyr Met Gly Val Val Leu Tyr Ala Pro Ser g Leu Ala Leu Asn Ala Val Thr Gly Phe Asp Leu Trp Leu Ser Val,.

Leu Ala Leu Gly Ile Val Cys Thr Val Tyr Thr Ala Leu G1y Gly Leu Lys Ala Val Ile Trp Thr Asp Val Phe Gln Thr Leu Val Met Phe Leu Gly Gln Leu Ala Val Ile Ile Val Gly Ser Ala Lys Val Gly Gly Leu Gly Arg Val Trp Ala Val Ala Ser Gln His Gly Arg Ile Ser Gly Phe Glu Leu Asp Pro Asp Pro Phe Val Arg His Thr Phe Trp Thr Leu Ala Phe Gly Gly Val Phe Met Met Leu Ser Leu Tyr Gly Val Asn Gln Ala Gln Val Gln Arg Tyr Leu Ser Ser Arg Thr Glu Lys Ala Ala Val Leu Ser Cys Tyr Ala Val Phe Pro Phe Gln Gln Val Ser Leu Cys Val Gly Cys Leu Ile Gly Leu Val Met Phe Ala Tyr Tyr Gln Glu Tyr Pro Met Ser Ile Gln Gln Ala Gln Ala Ala Pro Asp Gln Phe Val Leu Tyr Phe Val Met Asp Leu Leu Lys Gly Leu Pro Gly Leu Pro Gly Leu Phe Ile Ala Cys Leu Phe Ser Gly Ser Leu Ser Thr Ile Ser Ser Ala Phe Asn Ser Leu Ala Thr Val Thr Met Glu Asp Leu Ile Arg Pro Trp Phe Pro Glu Phe Ser Glu Ala Arg Ala Ile Met Leu Ser Arg Gly Leu Ala Phe Gly Tyr Gly Leu Leu Cys Leu Gly Met Ala Tyr Ile Ser Ser Gln Met Gly Pro Val Leu Gln Ala Ala Ile Ser Ile Phe Gly Met Val Gly Gly Pro Leu Leu Gly Leu Phe Cys Leu Gly Met Phe Phe Pro Cys Ala Asn Pro Pro Gly Ala Val Val Gly Leu Leu Ala Gly Leu Val Met Ala Phe Trp Ile Gly Ile Gly Ser Ile Val Thr Ser Met Gly Ser Ser Met Pro Pro Ser Pro Ser Asn Gly Ser Ser Phe Ser Leu Pro Thr Asn Leu Thr Val Ala Thr Val Thr Thr Leu Met Pro Leu Thr Thr Phe Ser Lys Pro Thr Gly Leu Gln Arg Phe Tyr Ser Leu Ser Tyr Leu Trp Tyr Ser Ala His Asn Ser Thr Thr Val Ile Val Val Gly Leu Ile Val Ser Leu Leu Thr Gly Arg Met Arg Gly Arg Ser Leu Asn Pro Ala Thr Ile Tyr Pro Val Leu Pro Lys Leu Leu Ser Leu Leu Pro Leu Ser Cys Gln Lys Arg Leu His Cys Arg Ser 575 580 58~.
Tyr Gly Gin Asp His Leu Asp Thr Gly Leu Phe Pro Glu Lys Pro Arg Asn Gly Val Leu Gly Asp Ser Arg Asp Lys Glu Ala Met Ala Leu Asp Gly Thr Ala Tyr Gln Gly Ser Ser Ser Thr Cys Ile Leu Gln Glu Thr Ser Leu <210> 6 <211> 535 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2414415CD1 <400> 6 Met Glu Glu Gly Ala Arg His Arg Asn Asn Thr Glu Lys Lys His Pro Gly Gly Gly Glu Ser Asp Ala Ser Pro Glu Ala Gly Ser Gly Gly Gly Gly Val Ala Leu Lys Lys Glu Ile Gly Leu Val Ser Ala Cys Gly Ile Ile Val Gly Asn Ile Ile Gly Ser Gly Ile Phe Val Ser Pro Lys Gly Val Leu Glu Asn Ala Gly Ser Val Gly Leu Ala Leu Ile Val Trp Ile Val Thr Gly Phe Ile Thr Val Val Gly Ala Leu Cys Tyr Ala Glu Leu Gly Val Thr Ile Pro Lys Ser Gly Gly Asp Tyr Ser Tyr Val Lys Asp Ile Phe Gly Gly Leu Ala Gly Phe Leu Arg Leu Trp Ile Ala Val Leu Val Ile Tyr Pro Thr Asn Gln Ala Val Ile Ala Leu Thr Phe Ser Asn Tyr Val Leu Gln Pro Leu Phe Pro Thr Cys Phe Pro Pro Glu Ser Gly Leu Arg Leu Leu Ala Ala Ile Cys Leu Leu Leu Leu Thr Trp Val Asn Cys Ser Ser Val Arg Trp Ala Thr Arg Val Gln Asp Ile Phe Thr Ala Gly Lys Leu Leu Ala Leu Ala Leu Ile Ile Ile Met Gly Ile Val Gln Ile Cys Lys Gly Glu Tyr Phe Trp Leu Glu Pro Lys Asn Ala Phe Glu Asn Phe Gln Glu Pro Asp Ile Gly Leu Val Ala Leu Ala Phe Leu Gln Gly Ser Phe Ala Tyr Gly Gly Trp Asn Phe Leu Asn Tyr Val Thr 245 250 , 255 Glu Glu Leu Val Asp Pro Tyr Lys Asn Leu Pro Arg Ala Ile Phe Ile Ser Ile Pro Leu Val Thr Phe Val Tyr Val Phe Ala Asn Val Ala Tyr Val Thr Ala Met Ser Pro Gln Glu Leu Leu Ala Ser Asn Ala Val Ala Val Thr Phe Gly Glu Lys Leu Leu Gly Val Met Ala Trp Ile Met Pro Ile Ser Val Ala Leu Ser Thr Phe Gly Gly Val Asn Gly Ser Leu Phe Thr Ser Ser Arg Leu Phe Phe Ala Gly Ala Arg Glu Gly His Leu Pro Ser Val Leu Ala Met Ile His Val Lys Arg Cys Thr Pro Ile Pro Ala Leu Leu Phe Thr Cys Ile Ser Thr Leu Leu Met Leu Val Thr Ser Asp Met Tyr Thr Leu Ile Asn Tyr Val Gly Phe Ile Asn Tyr Leu Phe Tyr Gly Val Thr Val Ala Gly Gln Ile Val Leu Arg Trp Lys Lys Pro Asp Ile Pro Arg Pro Ile Lys Ile Asn Leu Leu Phe Pro Ile Ile Tyr Leu Leu Phe Trp Ala Phe Leu Leu Val Phe Ser Leu Trp Ser Glu Pro Val Val Cys Gly Ile Gly Leu Ala Ile Met Leu Thr Gly Val Pro Val Tyr Phe Leu Gly Val Tyr Trp Gln His Lys Pro Lys Cys Phe Ser Asp Phe Ile Glu Leu Leu Thr Leu Val Ser Gln Lys Met Cys Val Val Val Tyr Pro Glu Val Glu Arg Gly Ser Gly Thr Glu Glu Ala Asn Glu Asp Met Glu Glu Gln Gln Gln Pro Met Tyr Gln Pro Thr Pro Thr Lys Asp Lys Asp Val Ala Gly Gln Pro Gln Pro <210> 7 <211> 456 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2466714CD1 <400> 7 Met Glu Ala Ser Trp Gly Ser Phe Asn Ala Glu Arg Gly Trp Tyr Val Ser Val Gln Gln Pro Glu Glu Ala Glu Ala Glu Glu Leu Ser Pro Leu Leu Ser Asn Glu Leu His Arg Gln Arg Ser Pro Gly Val Ser Phe Gly Leu Ser Val Phe Asn Leu Met Asn Ala Ile Met Gly Ser Gly Ile Leu Gly Leu Ala Tyr Val Met Ala Asn Thr Gly Val Phe Gly Phe Ser Phe Leu Leu Leu Thr Val Ala Leu Leu Ala Ser Tyr Ser Val His Leu Leu Leu Ser Met Cys Ile Gln Thr Ala Val Thr Ser Tyr Glu Asp Leu Gly Leu Phe Ala Phe Gly Leu Pro Gly Lys Leu Val Val Ala Gly Thr Ile Ile Ile Gln Asn Ile Gly Ala Met Ser Ser Tyr Leu Leu Ile Ile Lys Thr Glu Leu Pro Ala Ala Ile Ala Glu Phe Leu Thr Gly Asp Tyr Asn Arg Tyr Trp Tyr Leu Asp Gly Gln Thr Leu Leu Ile Ile Ile Cys Val Gly Ile Val Phe Pro Leu Ala Leu Leu Pro Lys Ile Gly Phe Leu Gly Tyr Thr Ser Ser Leu Ser Phe Phe Phe Met Met Phe Phe Ala Leu Val Val Ile Ile Lys Lys Trp Ser Ile Pro Cys Pro Leu Thr Leu Asn Tyr Val Glu Lys Gly Phe Gln Ile Ser Asn Val Thr Asp Asp Cys Lys Pro Lys Leu Phe His Phe Ser Lys Glu Ser Ala Tyr Ala Leu Pro Thr Met Ala Phe Sex Phe Leu Cys His Thr Ser Ile Leu Pro Ile Tyr Cys Glu Leu Gln Ser Pro Ser Lys Lys Arg Met Gln Asn Val Thr Asn Thr Ala Ile Ala Leu Ser Phe Leu Ile Tyr Phe Ile Ser Ala Leu Phe Gly Tyr Leu Thr Phe Tyr Asp Lys Val Glu Ser Glu Leu Leu Lys Gly Tyr Ser Lys Tyr Leu Ser His Asp Val Val Val Met Thr Val Lys Leu Cys Ile Leu Phe Ala Val Leu Leu Thr Val Pro Leu Ile His Phe Pro Ala Arg Lys Ala Val Thr Met Met Phe Phe Ser Asn Phe Pro Phe Ser Trp Ile Arg His Phe Leu Ile Thr Leu Ala Leu Asn Ile Ile Ile Val Leu Leu Ala Ile Tyr Val Pro Asp Ile Arg Asn Val Phe Gly Val Val Gly Ala Ser Thr Ser Thr Cys Leu Ile Phe Ile Phe Pro Gly Leu Phe Tyr Leu Lys Leu Ser Arg Glu Asp Phe Leu Ser Trp Lys Lys Leu Gly Ala Phe Val Leu Leu Ile Phe Gly Ile Leu Val Gly Asn Phe Ser Leu Ala Leu Ile Ile 12.

Phe Asp Trp Ile Asn Lys <210> 8 <211> 325 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2617942CD1 <400> 8 Met Phe Ala Asn Leu Lys Tyr Val Ser Leu Gly Ile Leu Val Phe Gln Thr Thr Ser Leu Val Leu Thr Met Arg Tyr Ser Arg Thr Leu Lys Glu Glu Gly Pro Arg Tyr Leu Ser Ser Thr Ala Val Val Val Ala Glu Leu Leu Lys Ile Met Ala Cys Ile Leu Leu Val Tyr Lys Asp Ser Lys Cys Ser Leu Arg Ala Leu Asn Arg Val Leu His Asp Glu Ile Leu Asn Lys Pro Met Glu Thr Leu Lys Leu Ala Ile Pro Ser Gly Ile Tyr Thr Leu Gln Asn Asn Leu Leu Tyr Val Ala Leu Ser Asn Leu Asp Ala Ala Thr Tyr Gln Val Thr Tyr Gln Leu Lys Ile Leu Thr Thr Ala Leu Phe Ser Val Ser Met Leu Ser Lys Lys Leu Gly Val Tyr Gln Trp Leu Ser Leu VaI Ile Leu Met Thr Gly Val Ala Phe Val Gln Trp Pro Ser Asp Ser Gln Leu Asp Ser Lys Glu Leu Ser Ala Gly Ser Gln Phe Val Gly Leu Met Ala Val Leu Thr Ala Cys Phe Ser Ser Gly Phe Ala Gly Val Tyr Phe Glu Lys Ile Leu Lys Glu Thr Lys Gln Ser Val Trp Ile Arg Asn Ile Gln Leu Gly Phe Phe Gly Ser Ile Phe Gly Leu Met Gly Val Tyr Ile Tyr Asp Gly Glu Leu Val Ser Lys Asn Gly Phe Phe Gln Gly Tyr Asn Arg Leu Thr Trp Ile Val Val Val Leu Gln Ala Leu Gly Gly Leu Val Ile Ala Ala Val Ile Lys Tyr Ala Asp Asn Ile Leu Lys Gly Phe Ala Thr Ser Leu Ser Ile Ile Leu Ser Thr Leu Ile Ser Tyr Phe Trp Leu Gln Asp Phe Val Pro Thr Ser Val Phe Phe Leu Gly Ala Ile Leu Val Ile Thr Ala Thr Phe Leu Tyr Gly Tyr Asp Pro Lys Pro Ala Gly Asn Pro Thr Lys Ala <210> 9 <211> 178 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2945431CD1 <400> 9 Met Ser Leu Ser Pro Arg Ser Gln Leu Ala Ile Ile Pro Gln Glu Pro Phe Leu Phe Ser Gly Thr Val Arg Glu Asn Leu Asp Pro Gln Gly Leu His Lys Asp Arg Ala Leu Trp Gln Ala Leu Lys Gln Cys His Leu Ser Glu Val Ile Thr Ser Met Gly Gly Leu Asp Gly Glu Leu Gly Glu Gly Gly Arg Ser Leu Ser Leu Gly Gln Arg Gln Leu Leu Cys Leu Ala Arg Ala Leu Leu Thr Asp Ala Lys Ile Leu Cys Ile Asp Glu Ala Thr Ala Ser Val Asp GIn Lys Thr Asp Gln Leu Leu Gln Gln Thr Ile Cys Lys Arg Phe Ala Asn Lys Thr Val Leu Thr Ile Ala His Arg Leu Asn Thr Ile Leu Asn Ser Asp Arg Val Leu Val Leu Gln Ala Gly Arg Val Val Glu Leu Asp Ser Pro Ala Thr Leu Arg Asn Gln Pro His Ser Leu Phe Gln GIn Leu Leu Gln Ser Ser Gln Gln Gly Val Pro Ala Ser Leu Gly Gly Pro <210> 10 <211> 255 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 4074113CD1 <400> 10 Met Glu Arg Glu Met Glu Gly Arg Pro Leu His Asn Glu Gly Trp Ile Asp Arg Ser Arg Val Gln Gln Lys Asp Leu Pro Asn Lys Cys Pro Gln Thr Leu Trp Ser Glu Gln Ala Phe Pro Pro Asn Pro Gly Gln Val Gly Ile Val Gly Arg Thr Gly Ala Gly Lys Ser Ser Leu Ala Ser Gly Leu Leu Arg Leu Pro Glu Ala Ala Glu Gly Gly Ile Trp Ile Asp Gly Val Pro Ile Ala His Val Gly Leu His Thr Leu Arg Ser Arg Ile Ser Ile Ile Pro Gln Asp Pro Ile Leu Phe Pro Gly Ser Leu Arg Met Asn Leu Asp Leu Leu Gln Glu His Ser Asp Glu Ala Ile Trp Ala Ala Leu Glu Thr Val Gln Leu Lys Ala Leu Val Ala Ser Leu Pro Gly Gln Leu Gln Tyr Lys Cys Ala Asp Arg Gly Glu Asp Leu Ser Val Gly Gln Lys Gln Leu Leu Cys Leu Ala Arg Ala Leu Leu Arg Lys Thr Gln Ile Leu Ile Leu Asp Glu Ala Thr Ala Ala Val Asp Pro Gly Thr Glu Leu Gln Met Gln Ala Met Leu Gly Ser Trp Phe Ala Gln Cys Thr Val Leu Leu Ile Ala His Arg Leu Arg Ser Val Met Asp Cys Ala Arg Val Leu Val Met Asp Lys Gly Gln Val Ala Glu Ser Gly Ser Pro Ala Gln Leu Leu Ala Gln Lys Gly Leu Phe Tyr Arg Leu Ala Gln Glu Ser Gly Leu Val <210> 11 <211> 462 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 1413743CD1 <400> 11 Met Ala Gln Val Ser Ile Asn Asn Asp Tyr Ser Glu Trp Asp Leu Ser Thr Asp Ala Gly Glu Arg Ala Arg Leu Leu Gln Ser Pro Cys Val Asp Thr Ala Pro Lys Ser Glu Trp Glu Ala Ser Pro Gly Gly Leu Asp Arg Gly Thr Thr Ser Thr Leu Gly Ala Ile Phe Ile Val Val Asn Ala Cys Leu Gly Ala Gly Leu Leu Asn Phe Pro Ala Ala Phe Ser Thr Ala Gly Gly Val Ala Ala Gly Ile Ala Leu Gln Met Gly Met Leu Val Phe Ile Ile Ser Gly Leu Val Ile Leu Ala Tyr Cys Ser Gln Ala Ser Asn Glu Arg Thr Tyr Gln Glu VaI Val Trp Ala Val Cys Gly Lys Leu Thr Gly Val Leu Cys Glu Val Ala Ile Ala Val Tyr Thr Phe Gly Thr Cys Ile Ala Phe Leu Ile Ile Ile Gly Asp Gln Gln Asp Lys Ile Ile Ala Val Met Ala Lys Glu Pro Glu Gly Ala Ser Gly Pro Trp Tyr Thr Asp Arg Lys Phe Thr Ile Ser Leu Thr Ala Phe Leu Phe Ile Leu Pro Leu Ser Ile Pro Arg Glu Ile Gly Phe Gln Lys Tyr Ala Ser Phe Leu Ser Val Val Gly Thr Trp Tyr Val Thr Ala Ile Val Ile Ile Lys Tyr Ile Trp Pro Asp Lys Glu Met Thr Pro Gly Asn Ile Leu Thr Arg Pro Ala Ser Trp Met Ala Val Phe Asn Ala Met Pro Thr Ile Cys Phe Gly Phe Gln Cys His Val Ser Ser Val Pro Val Phe Asn Ser Met Gln Gln Pro Glu Val Lys Thr Trp Gly Gly Val Val Thr Ala Ala Met Val Ile Ala Leu Ala Val Tyr Met Gly Thr Gly Ile Cys Gly Phe Leu Thr Phe GIy Ala Ala Val Asp Pro Asp Val Leu Leu Ser Tyr Pro Ser Glu Asp Met Ala Val Ala Val Ala Arg Ala Phe Ile Ile Leu Ser Val Leu Thr Ser Tyr Pro Ile Leu His Phe Cys Gly Arg Ala Val Val Glu Gly Leu Trp Leu Arg Tyr Gln Gly Val Pro Val Glu Glu Asp Val Gly Arg Glu Arg Arg Arg Arg Val Leu Gln Thr Leu Val Trp Phe Leu Leu Thr Leu Leu Leu Ala Leu Phe Ile Pro Asp Ile Gly Lys Val Ile Ser Val Ile Gly Gly Leu Ala Ala Cys Phe Ile Phe Val Phe Pro Gly Leu Cys Leu Ile Gln Ala Lys Leu Ser Glu Met Glu Glu Val Lys Pro Ala Ser Trp Trp Val Leu Val Ser Tyr Gly Val Leu Leu Val Thr Leu Gly Ala Phe Ile Phe Gly Gln Thr Thr Ala Asn Ala Ile Phe Val Asp Leu Leu Ala <210> 12 <211> 758 <212> PRT

<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 1733477CD1 <400> 12 Met Gly Leu Ala Asp Ala Ser Gly Pro Arg Asp Thr Gln Ala Leu Leu Ser Ala Thr Gln Ala Met Asp Leu Arg Arg Arg Asp Tyr His Met Glu Arg Pro Leu Leu Asn Gln Glu His Leu Glu Glu Leu Gly Arg Trp Gly Ser Ala Pro Arg Thr His Gln Trp Arg Thr Trp Leu Gln Cys Ser Arg Ala Arg Ala Tyr Ala Leu Leu Leu Gln His Leu Pro Val Leu Val Trp Leu Pro Arg Tyr Pro Val Arg Asp Trp Leu Leu Gly Asp Leu Leu Ser Gly Leu Ser Val Ala Ile Met Gln Leu Pro Gln Gly Leu Ala Tyr Ala Leu Leu Ala Gly Leu Pro Pro Val Phe Gly Leu Tyr Ser Ser Phe Tyr Pro Val Phe Ile Tyr Phe Leu Phe Gly Thr Ser Arg His Ile Ser Val Gly Thr Phe Ala Val Met Ser Val Met Val Gly Gly Val Thr Glu Ser Leu Ala Pro Gln Ala Leu Asn Asp Ser Met Ile Asn Glu Thr Ala Arg Asp Ala Ala Arg 170 1?5 180 Val Gln Val Ala Ser Thr Leu Ser Val Leu Val Gly Leu Phe Gln Val Gly Leu Gly Leu Ile His Phe Gly Phe Val Val Thr Tyr Leu Ser Glu Pro Leu Val Arg Gly Tyr Thr Thr Ala Ala Ala Val Gln Val Phe Val Ser Gln Leu Lys Tyr Val Phe Gly Leu His Leu Ser Ser His Ser Gly Pro Leu Ser Leu Ile Tyr Thr Val Leu Glu Val Cys Trp Lys Leu Pro Gln Ser Lys Val Gly Thr Val VaI Thr Ala Ala Val Ala Gly Val Val Leu Val Val Val Lys Leu Leu Asn Asp Lys Leu Gln Gln Gln Leu Pro Met Pro Ile Pro Gly Glu Leu Leu Thr Leu Ile Gly Ala Thr Gly Ile Ser Tyr Gly Met Gly Leu Lys His Arg Phe Glu Val Asp Val Val Gly Asn Ile Pro Ala Gly Leu Val Pro Pro Val Ala Pro Asn Thr Gln Leu Phe Ser Lys Leu Val Gly Ser Ala Phe Thr Ile Ala Val Val Gly Phe Ala Ile Ala Ile Ser Leu Gly Lys Ile Phe Ala Leu Arg His Gly Tyr Arg Val Asp 365 370 375_ Ser Asn Gln Glu Leu Val Ala Leu Gly Leu Ser Asn Leu Ile Gly Gly Ile Phe Gln Cys Phe Pro Val Ser Cys Ser Met Ser Arg Ser Leu Val Gln Glu Ser Thr Gly Gly Asn Sex Gln Val Ala Gly Ala Ile Ser Ser Leu Phe Ile Leu Leu Ile Ile Val Lys Leu Gly Glu Leu Phe His Asp Leu Pro Lys Ala Val Leu Ala Ala Ile Ile Ile Val Asn Leu Lys Gly Met Leu Arg Gln Leu Ser Asp Met Arg Ser Leu Trp Lys Ala Asn Arg Ala Asp Leu Leu Ile Trp Leu Val Thr Phe Thr Ala Thr Ile Leu Leu Asn Leu Asp Leu Gly Leu Val Val Ala Val Ile Phe Ser Leu Leu Leu Val Val Val Arg Thr Gln Met Pro His Tyr Ser Val Leu Gly Gln Val Pro Asp Thr Asp Ile Tyr Arg Asp Val Ala Glu Tyr Ser Glu Ala Lys Glu Val Arg Gly Val Lys VaI Phe Arg Ser Ser Ala Thr Val Tyr Phe Ala Asn Ala Glu Phe Tyr Ser Asp Ala Leu Lys Gln Arg Cys Gly Val Asp Val Asp Phe Leu Ile Ser Gln Lys Lys Lys Leu Leu Lys Lys Gln Glu Gln Leu Lys Leu Lys Gln Leu Gln Lys Glu Glu Lys Leu Arg Lys Gln Ala Ala Ser Pro Lys Gly Ala Ser Val Ser Ile Asn Val Asn Thr Ser Leu Glu Asp Met Arg Ser Asn Asn Val Glu Asp Cys Lys Met Met Val Ser Ser Gly Asp Lys Met Glu Asp Ala Thr Ala Asn Gly Gln Glu Asp Ser Lys Ala Pro Asp Gly Ser Thr Leu Lys Ala Leu Gly Leu Pro Gln Pro Asp Phe His Ser Leu Ile Leu Asp Leu Gly Ala Leu Ser Phe Val Asp Thr Val Cys Leu Lys Ser Leu Lys Asn Ile Phe His Asp Phe Arg Glu Ile Glu Val Glu Val Tyr Met Ala Ala Cys His Ser Pro Val Val Ser Gln Leu Glu Ala Gly His Phe Phe Asp Ala Ser Ile Thr Lys Lys His Leu Phe Ala Ser Val His Asp Ala Val Thr Phe Ala Leu Gln His Pro Arg Pro Val Pro Asp Ser Pro Val Ser Val Thr Arg Leu 1 g.

<220> 13 _ <211> 336 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2641908CD1 <400> 13 Met Met Gly Pro Gly Leu Ala Phe Gly Leu Gly Ser Leu Met Leu Arg Leu Tyr Val Asp Ile Asn Gln Met Pro Glu Gly Gly Ile Ser Leu Thr Ile Lys Asp Pro Arg Trp Val Gly Ala Trp Trp Leu Gly Phe Leu Ile Ala Ala Gly Ala Val Ala Leu Ala Ala Ile Pro Tyr Phe Phe Phe Pro Lys Glu Met Pro Lys Glu Lys Arg Glu Leu Gln Phe Arg Arg Lys Val Leu Ala Val Thr Asp Ser Pro Ala Arg Lys Gly Lys Asp Ser Pro Ser Lys Gln Ser Pro Gly Glu Ser Thr Lys Lys Gln Asp Gly Leu Val Gln Ile Ala Pro Asn Leu Thr Val Ile Gln Phe Ile Lys Val Phe Pro Arg Val Leu Leu Gln Thr Leu Arg His Pro IIe Phe Leu Leu Val Val Leu Ser Gln Val Cys Leu Ser Ser Met Ala Ala Gly Met Ala Thr Phe Leu Pro Lys Phe Leu Glu Arg Gln Phe Ser Ile Thr Ala Ser Tyr Ala Asn Leu Leu Ile Gly Cys Leu Ser Phe Pro Ser Val Ile Val Gly Ile Val Val Gly Gly Val Leu Val Lys Arg Leu His Leu Gly Pro Val Gly Cys Gly Ala Leu Cys Leu Leu Gly Met Leu Leu Cys Leu Phe Phe Ser Leu Pro Leu Phe Phe Ile Gly Cys Ser Ser His Gln Ile Ala Gly Ile Thr His Gln Thr Ser Ala His Pro Gly Leu Glu Leu Sex Pro Ser Cys Met Glu Ala Cys Ser Cys Pro Leu Asp Gly Phe Asn Pro Val Cys Asp Pro Ser Thr Arg Val Glu Tyr Ile Thr Pro Cys His Ala Gly Cys Ser Ser Trp Val Val Gln Asp Ala Leu Asp Asn Ser Gln Ser Pro Pro Thr Ser His Pro His Ala Gly His Gln His Leu Asn Leu Arg Leu Leu Gln Gly Glu Thr Trp Ala Ala Leu Ala Gly Ala Glu Glu Pro Val Asp Gly Ala <210> 14 <211> 103 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2656554CD1 <400> 14 Met Glu Arg Gln Ser Arg Val Met Ser Glu Lys Asp Glu Tyr Gln Phe Gln His Gln Gly Ala Val Glu Leu Leu Val Phe Asn Phe Leu Leu Ile Leu Thr Ile Leu Thr Ile Trp Leu Phe Lys Asn His Arg Phe Arg Phe Leu His Glu Thr Gly Gly Ala Met Val Tyr Asp Lys Pro Pro Lys Phe Ala Met Ser Arg Glu Gln Met Ser Gln Ser Cys Ser His Thr Ala His Asn Ala Ser Leu Leu Thr Asp Ala Gly Pro Leu Ser Cys Gly Glu Ser Arg Ala Ser Cys Leu Phe Leu <210> 15 <211> 123 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2719228CD1 <400> 15 Met Gln Gly Met Gly Leu Gly Leu Ser Ser Val Phe Ala Leu Cys Leu Gly His Thr Ser Ser Phe Cys Glu Ser Val Val Phe Ala Ser Ala Ser Ile Gly Leu Gln Thr Phe Asn His Ser Gly Ile Ser Val Asn Ile Gln Asp Leu Ala Pro Ser Cys Ala Gly Phe Leu Phe Gly Val Ala Asn Thr Ala Gly Ala Leu Ala Gly Val Val Gly Val Cys Leu Gly Gly Tyr Leu Met Glu Thr Thr Gly Ser Trp Thr Cys Leu Phe Asn Leu Val Ala Ile Ile Ser Asn Leu Gly Leu Cys Thr Phe Leu Val Phe Gly Gln Ala Gln Arg Val Asp Leu Ser Ser Thr His Glu Asp Leu <210> 16 ' <211> 222 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 3657824CD1 <400> 16 Met Lys Gln Glu Ser Ala Ala Pro Asn Thr Pro Pro Thr Ser Gln Ser Pro Thr Pro Ser Ala Gln Phe Pro Arg Asn Asp Gly Asp Pro Gln Ala Leu Trp Ile Phe Gly Tyr Gly Ser Leu Val Trp Arg Pro Asp Phe Ala Tyr Ser Asp Ser Arg Val Gly Phe Val Arg Gly Tyr Ser Arg Arg Phe Trp Gln Gly Asp Thr Phe His Arg Gly Ser Asp Lys Met Pro Gly Arg Val Val Thr Leu Leu Glu Asp His Glu Gly Cys Thr Trp Gly Val Ala Tyr Gln Val Gln Gly Glu Gln Val Ser Lys Ala Leu Lys Tyr Leu Asn Val Arg Glu Ala Val Leu Gly Gly 110 ~ 115 120 Tyr Asp Thr Lys Glu Val Thr Phe Tyr Pro Gln Asp Ala Pro Asp 125 130 . 135 Gln Pro Leu Lys Ala Leu Ala Tyr Val Ala Thr Pro Gln Asn Pro Gly Tyr Leu Gly Pro Ala Pro Glu Glu Ala Ile Ala Thr Gln Ile Leu Ala Cys Arg Gly Phe Ser Gly His Asn Leu Glu Tyr Leu Leu Arg Leu Ala Asp Phe Met Gln Leu Cys Gly Pro Gln Ala Gln Asp Glu His Leu Ala Ala Ile Val Asp Ala Val Gly Thr Met Leu Pro Cys Phe Cys Pro Thr Glu Gln Ala Leu Ala Leu Val <210> 17 <211> 111 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 5378485CD1 <400> 17 Met Leu Ser Ala Leu Pro Gly Trp Gly Pro Ala His Leu Gln Arg Pro Leu Leu Gly Pro Ala Ser Cys Leu Gly Ile Leu Arg Pro Ala 20 25 30' Met Thr Ala His Ser Phe Ala Leu Pro Val Ile Ile Phe Thr Thr Phe Trp Gly Leu Val Gly Ile Ala Gly Pro Trp Phe Val Pro Lys Gly Pro Asn Arg Gly Val Ile Ile Thr Met Leu Val Ala Thr Ala Val Cys Cys Tyr Leu Phe Trp Leu Ile Ala Ile Leu Ala Gln Leu Asn Pro Leu Phe Gly Pro Gln Leu Lys Asn Glu Thr Ile Trp Tyr Val Arg Phe Leu Trp Glu <210> 18 <211> 1303 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 961344CB1 <400> 18 cccacgcgtc cgcccacgcg tccgcaagat atggatgcta acaggcgata aactcgagac 60 agctacctgc attgccaaaa gttcacatct cgtgtctaga acacaagata ttcatatttt 120 cagacaggta accagtcggg gagaggcaca tttggagctg aatgcatttc gaaggaagca 180 tgattgtgca ctagtcatat ctggggactc tctggaggtt tgtctaaagt actacgagca 240 tgaatttgtg gagctggcct gccagtgccc tgccgtggtt tgctgccgct gctcacccac 300 ccagaaggcc cgcattgtga cactgctgca gcagcacaca gggagacgca cctgcgccat 360 cggtgatgga ggaaatgatg tcagcatgat tcaggcagca gactgtggga ttgggattga 420 gggaaaggag ggtaaacagg cctcgctggc ggccgacttc tccatcacgc agttccggca 480 cataggcagg ctgctcatgg tgcacgggcg gaacagctac aagaggtcgg cggcactcgg 540 ccagttcgtc atgcacaggg gccttatcat ctccaccatg caggctgtgt tttcctcagt 600 cttctacttc gcatccgtcc ctttgtatca gggcttcctc atggtggggt atgccaccat 660 atacaccatg ttcccagtgt tctccttagt gctggaccag gacgtgaagc cagagatggc 720 gatgctctac ccggagctgt acaaggacct caccaaggga agatccttgt ccttcaaaac 780 cttcctcatc tgggttttaa taagtattta ccaaggcggc atcctcatgt atggggccct 840 ggtgctcttc gagtctgagt tcgtccacgt ggtggccatc tccttcaccg cactgatcct 900 gaccgagctg ctgatggtgg cgctgaccgt ccgcacgtgg cactggctga tggtggtggc 960 cgagttcctc agcttaggct gctacgtgtc ctcactcgct tttctcaatg aatattttgg 1020 tataggcaga gtgtcttttg gagctttctt agatgttgcc tttatcacca ccgtgacctt 1080 cctgtggaaa gtgtcggcga tcaccgtggt cagctgcctc ccgctgtatg tcctcaagta 1140 cctgaggcgc aagctctctc ctcccagcta ctgcaagctg gcctcctaag gggctgtgca 1200 cccccagcgg gctggcccca gcaccttctg cccttcccag caccttgtgc ccttgccagt 1260 gaacgcaggg tttgccattg ctaccaagca agcaccacaa gaa 1303 <210> 19 <211> 3395 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 3128782CB1 <400> 19 cggaaatagc accgggcgcc gccacagtag ctgta~ctgc caccgcgatg ccgaaggcgc 60 ccaagcagca gccgccggag cccgagtgga tcggggacgg agagagcacg agcccatcag 120 acaaagtggt gaagaaaggg aagaaggaca agaagatcaa aaaaacgttc tttgaagagc 180 tggcagtaga agataaacag gctggggaag aagagaaagt gctcaaggag aaggagcagc 240 agcagcagca acagcaacag cagcagcaaa aaaaaaagcg agatacccga aaaggcaggc 300 ggaagaagga tgtggatgat gatggagaag agaaagagct catggagcgt cttaagaagc 360 tctcagtgcc aaccagtgat gaggaggatg aagtacccgc cccaaaaccc cgcggaggga 420 agaaaaccaa gggtggtaat gtttttgcag ccctgattca ggatcagagt gaggaagagg 480 aggaggaaga aaaacatcct cctaagcctg ccaagccgga gaagaatcgg atcaataagg 540 ccgtatctga ggaacagcag cctgcactca agggcaaaaa gggaaaggaa gagaagtcaa 600 aagggaaggc taagcctcaa aataaattcg ctgctctgga caatgaagag gaggataaag 660 aagaagaaat tataaaggaa aaggagcctc ccaaacaagg gaaggagaag gccaagaagg 720 cagagcaggg ttcagaggaa gaaggagaag gggaagaaga ggaggaggaa ggaggagagt 780 ctaaggcaga tgatccctat gctcatctta gcaaaaagga gaagaaaaag ctgaaaaaac 840 agatggagta tgagcgccaa gtggcttcat taaaagcagc caatgcagct gaaaatgact 900 tctccgtgtc ccaggcggag atgtcctccc gccaagccat gttagaaaat gcatctgaca 960 tcaagctgga gaagttcagc atctccgctc atggcaagga gctgttcgtc aatgcagacc 1020 tgtacattgt agccggccgc cgctacgggc tggtaggacc caatggcaag ggcaagacca 1080 cactcctcaa gcacattgcc aaccgagccc tgagcatccc tcccaacatt gatgtgttgc 1140 tgtgtgagca ggaggtggta gcagatgaga caccagcagt ccaggctgtt cttcgagctg 1200 acaccaagcg attgaagctg ctggaagagg agcggcggct tcagggacag ctggaacaag 1260 gggatgacac agctgctgag aggctagaga aggtgtatga ggaattgcgg gccactgggg 1320 cggcagctgc agaggccaaa gcacggcgga tcctggctgg cctgggcttt gaccctgaaa 1380 tgcagaatcg acccacacag aagttctcag ggggctggcg catgcgtgtc tccctggcca 1440 gggcactgtt catggagccc acactgctga tgctggatga gcccaccaac cacctggacc 1500 tcaacgctgt catctggctt aataactacc tccagggctg gcggaagacc ttgctgatcg 1560 tctcccatga ccagggcttc ttggatgatg tctgcactga tatcatccac ctcgatgccc 1620 agcggctcca ctactatagg ggcaattaca tgaccttcaa aaagatgtac cagcagaagc 1680 agaaagaact gctgaaacag tatgagaagc aagagaaaaa gctgaaggag ctgaaggcag 1740 gcgggaagtc caccaagcag gcggaaaaac aaacgaagga agccctgact cggaagcagc 1800 agaaatgccg acggaaaaac caagatgagg aatcccagga ggcccctgag ctcctgaagc 1860 gccctaagga gtacactgtg cgcttcactt ttccagaccc cccaccactc agccctccag 1920 tgctgggtct gcatggtgtg acattcggct accagggaca gaaaccactc tttaagaact 1980 tggattttgg catcgacatg gattcaagga tttgcattgt gggccctaat ggtgtgggga 2040 agagtacgct actcctgctg ctgactggca agctgacacc gacccatggg gaaatgagaa 2100 agaaccaccg gctgaaaatt ggcttcttca accagcagta tgeagagcag ctgcgcatgg 2160 aggagacgcc cactgagtac ctgcagcggg gcttcaacct gccctaccag gatgcccgca 2220 agtgcctggg ccgcttcggc ctggagagtc acgcccacac catccagatc tgcaaactct 2280 ctggtggtca gaaggcgcga gttgtgtttg ctgagctggc ctgtcgggaa cctgatgtcc 2340 tcatcttgga cgagccaacc aataacctgg acatagagtc tattgatgct ctaggggagg 2400 ccatcaatga atacaagggt gctgtgatcg ttgtcagcca tgatgcccga ctcatcacag 2460 aaaccaattg ccagctgtgg gtggtggagg agcagagtgt tagccaaatc gatggtgact 2520 ttgaagacta caagcgggag gtgttggagg ccctgggtga agtcatggtc agccggcccc 2580 gagagtgaag ctttccttcc cagaagtctc ccgagagaca tatttgtgtg gcctagaagt 2640 cctctgtggt ctcccctcct ctgaagactg cctctggcct gcagctgacc tggcaaccat 2700 tcaggcacat gaaggtggag tgtgaccttg atgtgaccgg gatcccactc tgattgcatc 2760 catttctctg aaagacttgt ttgttctgct tctcttcata taactgagct ggccttatcc 2820 ttggcatccc cctaaacaaa caagaggtga ccaccttatt gtgaggttcc atccagccaa 2880 gtttatgtgg cctattgtct caggactctc atcactcaga agcctgcctc tgatttaccc 2940 tacagcttca ggcccagctg ccccccagtc tttgggtggt gctgttcttt tctggtggat 3000 ttaatgctga ctcactggta caaacagctg ttgaagctca gagctggagg tgagcttctg 3060 aggcctttgc cattatccag cccaagattt ggtgcctgca gcctcttgtc tggt_tgagga 3120 cttggggcag gaaaggaatg ctgctgaact tgaatttccc tttacaaggg gaagaaataa 3180 aggaaaggag ttgctgccga cctgtcactg tttggagatt gatgggagtt ggaactgttc 3240 tcagtcttga tttgctttat tcagttttct agcagctttt aatagtcccc tcttccccac 3300 taaatggatc ttgtttacag tattactgac agtgtttact gtttaaggat cataggattc 3360 cttaacccca accattcccg caaggaataa gcaat 3395 <210> 20 <211> 2549 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 1720440CB1 <400> 20 acacgtccgc atgcaagacc atcaggcgcg atatctttgg cgcctggtcc tcctgaaggt 60 gaaggcaaag gtgcgacagt gcctgcagga gcggcggaca gtgcccattt tgtttgcctc 120 taccgttcgg cgccaccccg acaagacggc cctgatcttc gagggcacag atacccactg 180 gaccttccgc cagctggatg agtactcaag cagtgtagcc aacttcctgc aggcccgggg 240 ctgaccatcg gcgatgtggc tgccatcttc atggagaacc gcaatgagtt cgtgggccta 300 tggctgggca tggccaagct cggtgtggag gcagccctca tcaacaccaa cctgcggcgg 360 gatgctctgc tccactgcct caccacctcg cgcgcacggg cccttgtctt tggcagcgaa 420 atggcctcag ccatctgtga ggtccatgcc agcctggacc cctcgctcag cctcttctgc 480 tctggctcct gggagcccgg tgcggtgcct ccaagcacag aacacctgga ccctctgctg 540 aaagatgctc ccaagcacct tcccagttgc cctgacaagg gcttcacaga taaactgttc 600 tacatctaca catccggcac cacagggctg cccaaggccg ccatcgtggt gcacagcagg 660 tattaccgca tggctgccct ggtgtactat ggattccgca tgcggcccaa cgacatcgtc 720 tatgactgcc tccccctcta ccactcagca ggaaacatcg tgggaatcgg ccagtgcctg 780 ctgcatggca tgacggtggt gattcggaag aagttctcag cctcccggtt ctgggacgat 840 tgtatcaagt acaactgcac gattgtgcag tacattggtg aactgtgccg ctacctcctg 900 aaccagccac cgcgggaggc agaaaaccag caccaggttc gcatggcact aggcaatggc 960 ctccggcagt ccatctggac caacttttcc agccgcttcc acatacccca ggtggctgag 1020 ttctacgggg ccacagagtg caactgtagc ctgggcaact tcgacagcca ggtgggggcc 1080 tgtggtttca atagccgcat cctgtcctcc gtgtacccca tccggttggt acgtgtcaac 1140 gaggacacca tggagctgat ccgggggccc gacggcgtct gcattccctg ccagccaggt 1200 gagccgggcc agctggtggg ccgcatcatc cagaaagacc ccctgcgccg cttcgatggc 1260 tacctcaacc agggcgccaa caacaagaag attgccaagg atgtcttcaa gaagggggac 1320 caggcctacc ttactggtga tgtgctggtg atggacgagc tgggctacct gtacttccga 1380 gaccgcactg gggacacgtt ccgctggaaa ggtgagaacg tgtccaccac cgaggtggaa 1440 ggcacactca gccgcctgct ggacatggct gacgtggccg tgtatggtgt cgaggtgcca 1500 ggaaccgagg gccgggccgg aatggctgct gtggccagcc ccactggcaa ctgtgacctg 1560 gagcgctttg ctcaggtctt ggagaaggaa ctgcccctgt atgcgcgccc catcttcctg 1620 cgcctcctgc ctgagctgca caaaacagga acctacaagt tccagaagac agagctacgg 1680 aaggagggct ttgacccggc tattgtgaaa gacccgctgt tctatctaga tgcccagaag 1740 ggccgctacg tcccgctgga ccaagaggcc tacagccgca tccaggcagg cgaggagaag 1800 ctgtgattcc ccccatccct ctgagggccg gcggatgctg gatccggagc cccaggttcc 1860 gccccagagc ggtcctggac aaggccagac caaagcaagc agggcctggc acctccatcc 1920 tgaggtgctg cccctccatc caaaactgcc aagtgactca ttgccttccc aacccttcca 1980 gaggctttct gtgaaagtct catgtccaag ttccgtcttc tgggctgggc aggccctctg 2040 gttcccaggc tgagactgac gggttttctc aggatgatgt cttgggtgag ggtagggaga 2100 ggacaagggg tcaccgagcc cttcccagag agcagggagc ttataaatgg aaccagagca 2160 24, gaagtcccca gactcaggaa gtcaacagag tgggcaggga cagtggtagc atccatctgg 2220 tggccaaaga gaatcgtagc cccagagctg cccaagttca ctgggctcca cccc_cacctc 2280 caggagggga ggagaggacc tgacatctgt aggtggcccc tgatgcccca tctacagcag 2340 gaggtcagga ccacgcccct ggcctctccc cactccccca tcctcctccc tgggtggctg 2400 cctgattatc cctcaggcag ggcctctcag tccttgtggg tctgtgtcac ctccatctca 2460 gtcttggcct ggctatgagg ggaggaggaa tgggagaggg ggctcagggg ccaataaact 2520 ctgccttgag tcctcctaaa aaaaaaaaa 2549 <210> 21 <211> 2562 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2274290CB1 <400> 21 gcgggagcca acatagagcc ctcagtggga tgagggtgaa actgctattg ccggcggctc 60 ctgttttacc gcgtcagcat gctggtgcat ttatttcggg tcgggattcg gggtggccca 120 ttcccaggca ggctgctacc gcccctccgc ttccagacat tctcagctgt caggtactct 180 gatggctacc gcagctcctc cctcctccgg gccgtggccc acctgcggtc ccagctctgg 240 gcccacctcc ctcgagcccc cctagctccc agatggagcc cctctgcctg gtgctgggtt 300 gggggagccc tgctaggccc catggtactg agtaagcatc cccacctctg ccttgtggcc 360 ctgtgtgagg cagaagaggc ccctcctgcc agctccacac cccatgtcgt ggggtctcgc 420 tttaactgga agctcttctg gcagtttctg cacccccacc tgctggtcct gggggtagcc 480 gtcgtgctgg ccttgggtgc ggcactcgtg aatgtacaga tccccctgct cctgggccag 540 ctggtagagg tcgtggccaa gtacacaagg gaccacgtag ggagtttcat gactgagtcc 600 cagaatctca gcacccacct gcttatcctc tatggtgtcc agggactgct gaccttcggg 660 tacctggtgc tgctgtccca cgttggcgag cgcatggctg tggacatgcg gagggccctc 720 ttcagctccc tgctccgaca agacatcacc ttctttgacg ccaataagac agggcagctg 780 gtgagccgct tgacaactga cgtgcaggag tttaagtcat ccttcaagct tgtcatctcc 840 caggggctgc gaagctgcac ccaggtggca ggctgcctgg tgtccctgtc catgctgtcg 900 acacgcctca cgctgctgct gatggtggcc acaccagccc tgatgggagt gggcaccctg 960 atgggctcag gcctccgaaa attgtctcgc cagtgtcagg agcagatcgc cagggcaatg 1020 ggcgtagcag acgaggccct gggcaatgtg cggactgtgc gtgccttcgc catggagcaa 1080 cgggaagagg agcgctatgg ggcagagctg gaagcctgcc gctgccgggc agaggagctg 1140 ggccgcggca tcgccttgtt ccaagggctt tccaacatcg ccttcaactg catggtcttg 1200 ggtaccctat ttattggggg ctcccttgtg gccggacagc agctgacagg gggagacctc 1260 atgtccttcc tggtggcctc ccagacagtg caaaggtcca tggccaacct ctctgtcctg 1320 tttgggcagg tggtccgggg gctgagtgca ggtgcccggg tctttgagta catggccctg 1380 aacccctgca tcccactgtc tgggggctgc tgcgtcccca aagagcagct gcgtggctcc 1440 gttacatttc agaacgtctg cttcagctac ccctgccgcc ccggcttcga ggtgctgaaa 1500 gacttcaccc tgacgctgcc ccctggcaag atcgtggccc tcgtgggcca gtctggcgga 1560 ggaaagacca ccgtggcttc cctgctggag cgcttctacg accccacggc aggcgtggtg 1620 atgctggatg ggcgggacct gcgcaccctt gacccctcct ggctccgggg ccaggttgtc 1680 ggcttcatca gccaggagcc cgtcctgttt gggacgacca tcatggaaaa catccgcttt 1740 gggaagctgg aagcttccga tgaagaggtg tacacagccg cccgggaagc gaatgctcac 1800 gagttcatca ccagcttccc cgagggctac aacacggtcg tcggtgaacg gggcactacc 1860 ctgtctgggg gccagaagca gcgcctggcc atcgcccgag cccttatcaa gcagcccacg 1920 gtgctgatac tggatgaagc taccagcgcg ctggatgcag agtccgagcg ggttgtacag 1980 gaggccctgg accgggccag tgcaggccgc acggtgctgg taattgccca ccggctcagc 2040 actgtccgtg gggcccactg cattgtcgtc atggccgatg gccgtgtctg ggaggctggg 2100 acacatgaag agctcctgaa gaaaggcggg ctatacgccg agctcatccg gaggcaggcc 2160 ctggatgccc cgaggacagc ggcccccccg cccaaaaagc cagaaggccc caggagccac 2220 cagcacaagt cctgagaagg gccccctgag gtgtggtcgc tgccaagcat cagtgttagg 2280 ctc agcctggggg agcctactgg ggactgagcc cccaggaggg ccagcatgtg 2340 gctgggg gagagtcgct gcggct9ctc ctgctcacaa taaagccggg gccgagcagc tggcagggga 2400 ggccaatccc tccctcccct ccccagtcct gccggctgcc tccctcccac cagagtctgc 2460 cagagtcatt gggctgcaat gggcagagac agagttccac gagacacctc cactctattc 2520 tccctttgcc cagacccctc cagacctctc aagagacgtt ct 2562 <210> 22 <211> 2314 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2740029CB1 <400> 22 cgctggtctt catgcgccct agccctcttt cggggatact ggccgacccc ctcttccttt 60 tcccctttag tgaaggcctc ccccgtcgcc gcgcggcttc ccggagccga ctgcagactc 120 cctcagcccg gtgttccccg cgtccggacg ccgaggtcgc ggcttcgcag aaactcgggc 180 ccctccatcc gccctcagaa aagggagcga tgttgatctc aggaagcaca aagggacctt 240 cctagctctg actgaaccac ggagctcacc ttggacagta tcactccgtg gaggaagact 300 gtgagactgt ggctggaagc cagattgtag ccacacatcc gcccctgccc taccccagag 360 ccctggagca gcaactggct gcagatcaca gacacagtga ggatatgagt gtaggggtga 420 gcacctcagc ccctctttcc ccaacctcgg gcacaagcgt gggcatgtct accttctcca 480 tcatggacta tgtggtgttc gtcctgctgc tggttctctc tcttgccatt gggctctacc 540 atgcttgtcg tggctggggc cggcatactg ttggtgagct gctgatggcg gaccgcaaaa 600 tgggctgcct tccggtggca ctgtccctgc tggccacctt ccagtcagcc gtggccatcc 660 tgggtgtgcc gtcagagatc taccgatttg ggacccaata ttggttcctg ggctgctgct 720 actttctggg gctgctgata cctgcacaca tcttcatccc cgttttctac cgcctgcatc 780 tcaccagtgc ctatgagtac ctggagcttc gattcaataa aactgtgcga gtgtgtggaa 840 ctgtgacctt catctttcag atggtgatct acatgggagt tgtgctctat gctccgtcat 900 tggctctcaa tgcagtgact ggctttgatc tgtggctgtc cgtgctggcc ctgggcattg 960 tctgtaccgt ctatacagct ctgggtgggc tgaaggccgt catctggaca gatgtgttcc 1020 agacactggt catgttcctc gggcagctgg cagttatcat cgtggggtca gccaaggtgg 1080 gcggcttggg gcgtgtgtgg gccgtggctt cccagcacgg ccgcatctct gggtttgagc 1140 tggatccaga cccctttgtg cggcacacct tctggacctt ggccttcggg ggtgtcttca 1200 tgatgctctc cttatacggg gtgaaccagg ctcaggtgca gcggtacctc agttcccgca 1260 cggagaaggc tgctgtgctc tcctgttatg cagtgttccc cttccagcag gtgtccctct 1320 gcgtgggctg cctcattggc ctggtcatgt tcgcgtatta ccaggagtat cccatgagca 1380 ttcagcaggc tcaggcagcc ccagaccagt tcgtcctgta ctttgtgatg gatctcctga 1440 agggcctgcc aggcctgcca gggctcttca ttgcctgcct cttcagcggc tctctcagca 1500 ctatatcctc tgcttttaat tcattggcaa ctgttacgat ggaagacctg attcgacctt 1560 ggttccctga gttctctgaa gcccgggcca tcatgctttc cagaggcctt gcctttggct 1620 atgggctgct ttgtctagga atggcctata tttcctccca gatgggacct gtgctgcagg 1680 cagcaatcag catctttggc atggttgggg gaccgctgct gggactcttc tgccttggaa 1740 tgttctttcc atgtgctaac cctcctggtg ctgttgtggg cctgttggct gggctcgtca 1800 tggccttctg gattggcatc gggagcatcg tgaccagcat gggctccagc atgccaccct 1860 ctccctctaa tgggtccagc ttctccctgc ccaccaatct aaccgttgcc actgtgacca 1920 cactgatgcc cttgactacc ttctccaagc ccacagggct gcagcggttc tattccttgt 1980 cttacttatg gtacagtgct cacaactcca ccacagtgat tgtggtgggc ctgattgtca 2040 gtctactcac tgggagaatg cgaggccggt ccctgaaccc tgcaaccatt tacccagtgt 2100 tgccaaagct cctgtccctc cttccgttgt cctgtcagaa gcggctccac tgcaggagct 2160 acggccagga ccacctcgac actggcctgt ttcctgagaa gccgaggaat ggtgtgctgg 2220 gggacagcag agacaaggag gccatggccc tggatggcac agcctatcag gggagcagct 2280 ccacctgcat cctccaggag acctccctgt gatg 2314 <210> 23 <211> 2155 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2414415CB1 <400> 23 gtggttccta tttcggaaaa ggacgttcta attcaaagct ctctcccaat atatttacac 60 gaatacgcat ttagaaaggg aggcagcttt tgaggttgca atcctactga gaaggatgga 120 agaaggagcc aggcaccgaa acaacaccga aaagaaacac ccaggtgggg gcgagtcgga 180 cgccagcccc gaggctggtt ccggaggggg cggagtagcc ctgaagaaag agatcggatt 240 ggtcagtgcc tgtggtatca tcgtagggaa catcatcggc tctggaatct ttgtctcgcc 300 aaagggagtg ctggagaatg ctggttctgt gggccttgct ctcatcgtct ggattgtgac 360 gggcttcatc acagttgtgg gagccctctg ctatgctgaa ctcggggtca ccatccccaa 420 atctggaggt gactactcct atgtcaagga catcttcgga ggactggctg ggttcctgag 480 gctgtggatt gctgtgctgg tgatctaccc caccaaccag gctgtcatcg ccctcacctt 540 ctccaactac gtgctgcagc cgctcttccc cacctgcttc cccccagagt ctggccttcg 600 gctcctggct gccatctgct tattgctcct cacatgggtc aactgttcca gtgtgcggtg 660 ggccacccgg gttcaagaca tcttcacagc tgggaagctc ctggccttgg ccctgattat 720 catcatgggg attgtacaga tatgcaaagg agagtacttc tggctggagc caaagaatgc 780 atttgagaat ttccaggaac ctgacatcgg cctcgtcgca ctggctttcc ttcagggctc 840 ctttgcctat ggaggctgga actttctgaa ttacgtgact gaggagcttg ttgatcccta 900 caagaacctt cccagagcca tcttcatctc catcccactg gtcacatttg tgtatgtctt 960 tgccaatgtc gcttatgtca ctgcaatgtc cccccaggag ctgctggcat ccaacgccgt 1020 cgctgtgact tttggagaga agctcctagg agtcatggcc tggatcatgc ccatttctgt 1080 tgccctgtcc acatttggag gagttaatgg gtctctcttc acctcctctc ggctgttctt 1140 cgctggagcc cgagagggcc accttcccag tgtgttggcc atgatccacg tgaagcgctg 1200 caccccaatc ccagccctgc tcttcacatg catctccacc ctgctgatgc tggtcaccag 1260 cgacatgtac acactcatca actacgtggg cttcatcaac tacctcttct atggggtcac 1320 ggttgctgga cagatagtcc ttcgctggaa gaagcctgat atcccccgcc ccatcaagat 1380 caacctgctg ttccccatca tctacttgct gttctgggcc ttcctgctgg tcttcagcct 1440 gtggtcagag ccggtggtgt gtggcattgg cctggccatc atgctgacag gagtgcctgt 1500 ctatttcctg ggtgtttact ggcaacacaa gcccaagtgt ttcagtgact tcattgagct 1560 gctaaccctg gtgagccaga agatgtgtgt ggtcgtgtac cccgaggtgg agcggggctc 1620 agggacagag gaggctaatg aggacatgga ggagcagcag cagcccatgt accaacccac 1680 tcccacgaag gacaaggacg tggcggggca gccccagccc tgaggaccac cattccctgg 1740 ctactctctc cttcctcccc cttttatcct acctccctgc cttggtcccg ccaacacatg 1800 cgagtacaca cacacccctc tctctgcttt tgtcaggcag tggtaggact ttggtgtggg 1860 tggtgagaaa ttgtaaacaa aaactgacat tcatacccaa agaaccagcc tctcacccca 1920 gggtccatgt cccaggcccc actccagtgc tgcccacact cccagctgct ggaggagagg 1980 ggagatgcca aggtgccctg caggacctcc ctccgggcca caccctcagc tgcctcttca 2040 ggaaccggag ctcattactg ccttccctcc cagggaggcc ccttcagaga ggagaggcca 2100 caggagctgc attgtggggg gacaggctca agcaattctg tccccatcaa ggggt 2155 <210> 24 <2I1> 1475 pCT/US99/26048 <212> DNA
<213> Homo sapiens ' <220>
<221> misc_feature <223> Incyte ID No: 2466714CB1 <400> 24 ggagcgcagg gcaggggtag aggctcgtag atggaactgg tagtcagctg gagagcagca 60 tggaggcgtc ctgggggagc ttcaacgctg agcggggctg gtatgtctct gtccagcagc 120 ctgaagaagc ggaggccgaa gagttgagtc cgttgctaag caacgaactt cacagacagc 180 gatccccagg tgtttcattt ggtttatcag tgtttaattt gatgaatgcc atcatgggaa 240 gtggcatcct tggcttagct tatgttatgg ctaataccgg tgtctttgga tttagcttct 300 tgctgctgac agttgctctc ctggcttctt actcagtcca tcttctgctt agtatgtgta 360 ttcagacagc tgtaacatct tatgaagatc ttggactctt tgcatttgga ttacctggaa 420 agttggtggt ggcaggcacc ataataattc agaatattgg agctatgtca tcttatcttt 480 taattattaa aacagagctt cctgctgcta ttgcagaatt tttgactgga gactataata 540 gatattggta tcttgatgga caaacactac taataatcat atgtgttggc attgtgttcc 600 ctcttgcact tcttcccaaa ataggctttc ttggctacac aagtagttta tcatttttct 660 ttatgatgtt ctttgctctt gtggtaataa ttaaaaaatg gtccatccct tgtcctctga 720 cattaaatta tgtagagaaa ggcttccaga tttcaaatgt tacagatgat tgtaagccaa 780 agctctttca tttctccaaa gagagtgctt atgccttacc aaccatggct ttttcatttc 840 tctgccatac ctcaatattg cccatatact gtgaacttca aagtccttca aagaaaagaa 900 tgcagaatgt taccaataca gcaattgctt t.aagttttct catttatttt atatctgcac 960 tctttgggta cctcactttt tatgacaaag tggagtcaga attactaaaa ggttatagta 1020 aatacttatc acatgatgtt gttgtcatga ctgtgaagtt atgcatacta tttgctgtgc 1080 ttttgacagt ccctctaatc cacttccctg ccagaaaagc tgtaacaatg atgtttttct 1140 ccaattttcc attctcatgg attcgccatt ttttgatcac tctagcactc aatattatca 1200 tcgttttact tgcaatatat gttcctgaca ttagaaatgt atttggtgta gttggtgcca 1260 gtacatcaac atgtttgatt tttatattcc caggactatt ttatcttaaa cttagcagag 1320 aggattttct gtcatggaaa aagcttgggg cattcgtttt gctcatcttt ggaattttgg 1380 ttgggaattt tagtttagca ctcatcattt ttgattggat taataaataa aagaaatatt 1440 ttcctacttc ttacaagaat aataaaaaaa aaaaa 1475 <210> 25 <211> 1793 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2617942CB1 <400> 25 gcggtggcta cggcgacggg agccggcggc gctgcgggtc agcggtcgcg taggacccag 60 cggactcggc agcctggggc gcccggcgga gctgaaccgc ggcccccggt ggtgggctca 120 gccggtcgag ctgcgcggga ggcaaatgaa gataaaacaa tgttcgccaa cctaaaatac 180 gtttccctgg gaattttggt ctttcagact accagtttgg ttctaacaat gcgttattcc 240 agaactttaa aagaagaagg acctcgttat ctatcttcta cagcagtggt tgttgctgaa 300 cttttgaaga taatggcctg cattttattg gtctacaaag acagcaaatg tagtctaaga 360 gcactgaatc gagtactaca tgatgaaatt cttaataaac ctatggaaac acttaaactt 420 gctattccat cagggatcta tactcttcag aataatttac tgtatgtggc actatcaaat 480 ctagatgcag ctacttatca ggtcacgtat cagttaaaaa ttcttacaac agcattattt 540 tctgtgtcta tgcttagtaa aaaattgggt gtataccagt ggctgtccct agtaattttg 600 pCT/US99/26048 atgacaggag ttgcttttgt acagtggccc tcagattctc agcttgattc taaggaactt 660 tcagctggtt ctcaatttgt aggactcatg gcagttctca cagcatgttt ttcaagtggc 720 tttgctgggg tttactttga gaaaatctta aaagaaacaa aacaatcagt gtggataaga 780 aatattcagc ttggtttctt tggaagtata tttggattaa tgggtgtata catttatgat 840 ggagaactgg tatcaaagaa tggatttttt cagggatata accgactgac ctggatagta 900 gttgttcttc aggcacttgg aggccttgta atagctgctg ttattaagta tgcagataat 960 attttaaaag gatttgcaac ctctttatcg ataatattat caacattgat ctcctatttt 1020 tggcttcaag attttgtgcc aaccagtgtc tttttccttg gagccatcct tgtaataaca 1080 gctacttttt tgtatggtta tgatcccaaa cctgcaggaa atcccactaa agcatagttg 1140 tatactatct ttaactggtt tttcacgatg gggcactagg aatctcgaca ttaatcttgc 1200 acagaggact tctacagagt ctgagaagat atcatcatgc tgaatctgat catactgttt 1260 tttaaaagtt taaggataag acatgtgtat atgtaacaaa acacattgca tctagaaatc 1320 aaaacttgaa agtatttcca gggattagga ttagaaggaa tattagagga aacttgaaat 1380 ctgagtttaa aaagatttta cctttttgat tgctgcagaa atgtcctatg cactctttgc 1440 aagagcacac aacaaatgtc agataccaat ttttgcaaat tagatttaat cttattaaat 1500 gtttttatct tactctttct gtacagatat atcaaatcac atgaaatatt taaagtttga 1560 aaattataat tacctataaa gctgtgaaaa atagaagtat aatttgaaaa aacatttcac 1620 ttatcagaga tttttatatt tatacaaaag attacttaat gaaggattgc taaatgtttt 1680 tgggtcaatt accttaagat taatattccg ggtctgatct gtcagggaat aaatatcaaa 1740 tctaaatttt aatgtggggg ttcatactat ttctcccata agaattttag ggt 1793 <210> 26 <211> 1141 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2945431CB1 <400> 26 ctggtttgtg tgtgtgcacc tccgtgaaat gtaggcacct tgaggacaga gtccagcctt 60 tggtttcttt ggtattgctc atagcactgg cacagttcta ggtacccagc tactaacaga 120 tcatttggtg gggatggggt ggggagcaga gtggggttat gttcaggtct catacccagg 180 ctttcatgga ggtgctagcc ctgtagtcag aaactgagct gggagcagaa gtggctacat 240 ctccaaccac tagactccat gtcattgtcc cccagatccc agttggctat catcccccag 300 gagccctttt tgttcagtgg gactgttcgg gaaaacctgg acccccaggg cctacataag 360 gacagggcct tgtggcaggc cctgaagcag tgccacctga gtgaggtgat tacatccatg 420 ggtggtctgg atggtgagct gggtgagggg ggccggagct tatctcttgg gcagaggcag 480 ctgttgtgtt tggccagggc tctcctcaca gatgccaaga tcctgtgtat cgatgaggcc 540 acagcaagtg tggaccagaa gacagaccag ctgctccagc agaccatctg caaacgcttt 600 gccaacaaga cagtgctgac cattgcccat aggctcaaca cgatcctgaa ctcagaccgg 660 gtgctggtgc tacaagcggg gagagtggta gagctggact ccccggccac cctgcgcaac 720 cagccccact ccctgttcca gcagctgctg cagagcagcc agcagggagt ccctgcctca 780 ctcggaggtc cctgagccca atcccacacc ctgcagagtt ctcccctctc tctgatccag 840 gccgggccta tacagaggtg ctggctgctt gtttacattc tcctctgggg ctctacctct 900 ccacacttcc ccagaaggga aaagggcacc ctggattact ctttggaaat cactccttgg 960 tgggcagcat cctgaggctt ccccagaacc aggcctctgc tctggccctc ttgcatctgg 1020 aacgccaggt gggtttttct ggcataggag cccacttgca ttttcatagt tttatttgat 1080 aaaattccat cttacattct gtgtattaaa aaaataatat ttctggtgtg agaaaaaaaa 1140 a <210> 27 <211> 1371 .
<212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 4074113CB1 <400> 27 gactggatgg attgatgggt ggatatatag aaaggtagac agatggaaag agagatggaa 60 ggtagacctt tacacaatga gggatggata gacagatctc gggtacagca gaaagatctc 120 cccaataaat gcccacaaac cctctggtca gagcaggcct ttcctcccaa ccccgggcag 180 gtgggcatcg ttggcaggac cggggcaggg aagtcctccc tggccagtgg gctgctgcgg 240 ctcccagagg cagctgaggg tgggatctgg atcgacgggg tccccattgc ccacgtgggg 300 ctgcacacac tgcgctccag gatcagcatc atcccccagg accccatcct gttccctggc 360 tctctgcgga tgaacctcga cctgctgcag gagcactcgg acgaggctat ctgggcagcc 420 ctggagacgg tgcagctcaa agccttggtg gccagcctgc ccggccagct gcagtacaag 480 tgtgctgacc gaggcgagga cctgagcgtg ggccagaaac agctcctgtg tctggcacgt 540 gcccttctcc ggaagaccca gatcctcatc ctggacgagg ctactgctgc cgtggaccct 600 ggcacggagc tgcagatgca ggccatgctc gggagctggt ttgcacagtg cactgtgctg 660 ctcattgccc accgcctgcg ctccgtgatg gactgtgccc gggttctggt catggacaag 720 gggcaggtgg cagagagcgg cagcccggcc cagctgctgg cccagaaggg cctgttttac 780 agactggccc aggagtcagg cctggtctga gccaggaccc tcaaccgtac cccagttgga 840 ccagcccgca cagcctgcag tgctggagat ggaagtgacc cgtggtcatc gatagctcca 900 cacgatattg agtctagacc tgtgtttgct ctctgggagg aaaatggcag agaaagtggc 960 caattatcac agagcatcag agccggaagg acctagcaat acacaggtct gcccgggcag 1020 ggcccatctc gccctgtcca ccctgcagcc aatgtcaaca gcgactctca gccccgctgt 1080 actctggact cacctggggg cctcaagcac atgcccaggc tcccggctag acccttaaat 1140 cagaatctct gaggctggga actgccatgc tgtgtgtact ttttacaaat taacactttt 1200 attttgggat aatcccagac tcacatgcag ttaaagaaac aataatatag agagattcgt 1260 gtacttggta ccccatttca cccaatggta acatcttgca aaactctagg ataaagcatc 1320 acagccaggg tgttgacatt gacacaacaa tcttgctcgg atgtccgcga g 1371 <210> 28 <211> 2752 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 1413743CB1 <400> 28 gggaaagaat cccccaagct ccatttcatg agtaagcgtg agagccgctc agtttcctcc 60 agctctgctg aagccagcac agaagtagcc caaactcttc cctctgctga cagcaaattt 120 taggcaaagt cttgagaaag aagaaattgg gtccagaaag ggaagtgagg agaatcagat 180 cccagacctt tggggagaag gagcaaccgc ctctggcaca gcccatcagg gagaaagagc 240 aggttgagaa gagtcctaag ctaacagccc caaacaggtg ggtgttgctc agctccctga 300 ggcatgtggt tgtaaggcag aacccacaga ccttgcagga agaaggctct cggggccatg 360 gcccaggtca gcatcaacaa tgactacagc gagtgggact tgagcacgga tgccggggag 420 cgggctcggc tgctgcagag tccctgtgtg gacacagccc ccaagagtga gtgggaagcc 480 tctcctgggg gtctggacag aggcaccact tccacacttg gggccatctt catcgtcgtc 540 aacgcgtgcc tgggtgcagg gttactcaac ttcccagcag ccttcagcac tgcggggggc 600 gtggcagcag gcatcgcact gcagatgggt atgctggttt tcatcatcag tggccttgtc 660 atcctggcct actgctccca ggccagcaat gagaggacct accaggaggt ggtatgggct 720 gtgtgtggca agctgacagg tgtgctatgt gaggtggcca tcgctgtcta cacctttggc 780 acctgcattg ccttcctaat catcattggc gaccagcagg acaagattat agctgtgatg 840 gcgaaagagc cggagggggc cagcggccct tggtacacag accgcaagtt caccatcagc 900 ctcactgcct tcctcttcat cctgcccctc tccatcccca gggagattgg tttccagaaa 960 tatgccagct tcctgagcgt cgtgggtacc tggtacgtca cagccatcgt tatcatcaag 1020 tacatctggc cagataaaga gatgacccca gggaacatcc tgaccaggcc ggcttcctgg 1080 atggctgtgt tcaatgccat gcccaccatc tgcttcggat ttcagtgcca cgtcagcagt 1140 gtgcccgtct tcaacagcat gcagcagcct gaagtgaaga cctggggtgg agtggtgaca 1200 gctgccatgg tcatagccct cgctgtctac atggggacag gcatctgtgg cttcctgacc 1260 tttggagctg ctgtggatcc tgacgtgctc ctgtcctatc cctcggagga catggccgtg 1320 gccgttgccc gagccttcat catcctgagc gtgctcacct cctaccctat cctgcacttc 1380 tgtgggcggg cggtggtgga aggcctgtgg ctgcgctacc agggg9tgcc agtggaggag 1440 gacgtggggc gggagcggcg gcggcgagtg ctgcagacgc tggtctggtt cctgctcacc 1500 ctgctgctgg cgctcttcat ccctgacatc ggcaaggtga tctcagtcat tggaggcctg 1560 gccgcctgct tcatcttcgt cttcccaggg ctgtgcctca ttcaagccaa actctctgag 1620 atggaagagg tcaaaccagc cagctggtgg gtgctggtca gctacggagt cctcttggtc 1680 accctgggag ccttcatctt cggccagacc acagccaacg ccatctttgt ggatctcttg 1740 gcataaccac tgcctcccag ggaacacaag gcctttgcca ttggtcgcag gaacccatct 1800 cttagagcta tggggccatt cttagtccac gatcattcca actggtggga tgacatccgg 1860 acatcctctt ccagggactg gggcaaactc aggccccaca cctctggaca gctcaaatcc 1920 agtccccttc ctgctcccca gtcctggcag tgccgtggat ggcggcagga agtctcacat 1980 catagaggac ccctcctcct ctcccagttc tcaacttctc catgcctgga atccacgggt 2040 gaagagagtc ggtagatctc ataagaaaga atccagtctg acttccctct ggagaatgac 2100 tatggacaga aggccaccat cctccacaga gcaccctgtc ctgagtaggg gttgtgctca 2160 ttaccccagg ccagtggtag cttcctcagg agcctggcca cttccaacgg tagcactgaa 2220 gtcatgcaaa tgcatagtca ggtagattca gaccttgtcc acaccttcct gggcaacccc 2280 caccatgaac ctgtcagcct ctttcccata gctaatagac atttcccagg ccttgagggg 2340 ccccaccctg tctctttcat caaacctgat ggtccaggct gggcatccct ctcctcctcc 2400 atccccagac atcaccaggt ctaatgttta caaacggtgc cagcccggct ctgaagccaa 2460 gggccgtccc gtgccacggt gctgtgagta ttcctccgtt agctttcccc ataaggttgg 2520 gagtatctgc ttttgtgtct gagatgggcc cctcttttca gaggccgcag ggtgggtgat 2580 ggagaaggct gagaaccttt cagaccctct gtgtgggctg ggctggtcag aatcagggtg 2640 tacctccccg acaccttctt tttcagtgat gttttctctt ctccctgcct ttcctctgcc 2700 tcctcccctg ccagccctag cgtgactacc cagagacaaa aaaaaaaaaa as 2752 <210> 29 <211> 2580 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 1733477CB1 <400> 29 ggagcagccc gcaccggaca acttgcgagc catggggctg gcggatgcgt cgggaccgag 60 ggacacacag gcactgctgt ctgcaacaca agcaatggac ctgcggaggc gagactacca 120 catggaacgg ccgctgctga accaggagca tttggaggag ctggggcgct ggggctcagc 180 acctaggacc caccagtggc ggacctggtt gcagtgctcc cgtgctcggg cctatgccct 240 tctgctccaa cacctcccgg ttttggtctg gttaccccgg tatcctgtgc gtgactggct 300 cctgggtgac ctgttatccg gcctgagtgt ggccatcatg cagcttccgc agggcttggc 360 ctacgccctc ctggctggat tgccccccgt gtttggcctc tatagctcct tctaccctgt 420 cttcatctac ttcctgtttg gcacttcccg gcacatctcc gtggggacct ttgctgtcat 480 gtctgtgatg gtgggcggtg tgacagaatc cctggccccg caggccttga acgactccat 540 gatcaatgag acagccagag atgctgcccg ggtacaggtg gcctccacac tcagtgtcct 600 ggttggcctc ttccaggtgg ggctgggcct gatccacttc ggcttcgtgg tcacctacct 660 gtcagaacct cttgtccgag gctataccac agctgcagct gtgcaggtct tcgtctcaca 720 gctcaagtat gtgtttggcc tccatctgag cagccactct gggccactgt ccctcatcta 780 tacagtgctg gaggtctgct ggaagctgcc ccagagcaag gttggcaccg tggtcactgc 840 agctgtggct g9ggtggtgc tcgtggtggt gaagctgttg aatgacaagc tgcagcagca 900 gctgcccatg ccgatacccg gggagctgct cacgctcatc ggggccacag gcatctccta 960 tggcatgggt ctaaagcaca gatttgaggt agatgtcgtg ggcaacatcc ctgcagggct 1020 ggtgccccca gtggccccca acacccagct gttctcaaag ctcgtgggca gcgccttcac 1080 catcgctgtg gttgggtttg ccattgccat ctcactgggg aagatcttcg ccctgaggca 1140 cggctaccgg gtggacagca accaggagct ggtggccctg ggcctcagta accttatcgg 1200 aggcatcttc cagtgcttcc ccgtgagttg ctctatgtct cggagcctgg tacaggagag 1260 caccgggggc aactcgcagg ttgctggagc catctcttcc cttttcatcc tcctcatcat 1320 tgtcaaactt ggggaactct tccatgacct gcccaaggcg gtcctggcag ccatcatcat 1380 tgtgaacctg aagggcatgc tgaggcagct cagcgacatg cgctccctct ggaaggccaa 1440 tcgggcggat ctgcttatct ggctggtgac cttcacggcc accatcttgc tgaacctgga 1500 ccttggcttg gtggttgcgg tcatcttctc cctgctgctc gtggtggtcc ggacacagat 1560 gccccactac tctgtcctgg ggcaggtgcc agacacggat atttacagag atgtggcaga 1620 gtactcagag gccaaggaag tccggggggt gaaggtcttc cgctcctcgg ccaccgtgta 1680 ctttgccaat gctgagttct acagtgatgc gctgaagcag aggtgtggtg tggatgtcga 1740 cttcctcatc tcccagaaga agaaactgct caagaagcag gagcagctga agctgaagca 1800 actgcagaaa gaggagaagc ttcggaaaca ggctgcctcc cccaagggcg cctcagtttc 1860 cattaatgtc aacaccagcc ttgaagacat gaggagcaac aacgttgagg actgcaagat 1920 gatggtgagc tcaggagata agatggaaga tgcaacagcc aatggtcaag aagactccaa 1980 ggccccagat gggtccacac tgaaggccct gggcctgcct cagccagact tccacagcct 2040 catcctggac ctgggtgccc tctcctttgt ggacactgtg tgcctcaaga gcctgaagaa 2100 tattttccat gacttccggg agattgaggt ggaggtgtac atggcggcct gccacagccc 2160 tgtggtcagc cagcttgagg ctgggcactt cttcgatgca tccatcacca agaagcatct 2220 ctttgcctct gtccatgatg ctgtcacctt tgccctccaa cacccgaggc ctgtccccga 2280 cagccctgtt tcggtcacca gactctgaac atgctacatc ctgcccaaga ctgcacctct 2340 ggaggtgcag ggcacccttg agaagcccct cacccctagg ccgcctccag gtgctaccca 2400 ggagtcccct ccatgtacac acacacaact cagggaagga ggtcctggga ctccaagttc 2460 agcgctccag gtctgggaca gggcctgcat gcagtcaggc tggcagtggc gcggtacagg 2520 gagggaactg gtgcatattt tagcctcagg aataaagatt tgtctgctca aaaaaaaaaa 2580 <210> 30 <211> 1481 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2641908CB1 <400> 30 tgatgattgt gctggtggtg gtgatcatga cagagacaac aataacaatc atcacatcgt 60 gatggtaatg tcgtgactaa atttgtcatt tagtcacaac gatatgggtg atgtgaatga 120 gggtgatatt taagctgaaa ggaatagaaa tgatgatgac agcaactcgc ccctctacct 180 cgggatcctg tttgcagtga ccatgatggg gccaggcctg gcctttgggc tgggcagcct 240 catgctgcgc ctttatgtgg acattaacca gatgccagaa ggtggtatca gcctgaccat 300 aaaggacccc cgatgggtgg gtgcctggtg gctgggtttc ctcatcgctg ccggtgcagt 360 ggccctggct gccatcccct acttcttctt ccccaaggaa atgcccaagg aaaaacgtga 420 gcttcagttt cggcgaaagg tcttagcagt cacagactca cctgccagga agggcaagga 480 ctctccctct aagcagagcc ctggggagtc cacgaagaag caggatggcc tagtccagat 540 tgcaccaaac ctgactgtga tccagttcat taaagtcttc cccagggtgc tgctgcagac 600 cctacgccac cccatcttcc tgctggtggt cctgtcccag gtatgcttgt catccatggc 660 tgcgggcatg gccaccttcc tgcccaagtt cctggagcgc cagttttcca tcacagcctc 720 ctacgccaac ctgctcatcg gctgcctctc cttcccttcg gtcatcgtgg gcatcgtggt 780 gggtggcgtc ctggtcaagc ggctccacct gggccctgtg ggatgcggtg ccctttgcct 840 gctggggatg ctgctgtgcc tcttcttcag cctgccgctc ttctttatcg gctgctccag 900 ccaccagatt gcgggcatca cacaccagac cagtgcccac cctgggctgg agctgtctcc 960 aagctgcatg gaggcctgct cctgcccatt ggacggcttt aaccctgtct gcgaccccag 1020 cactcgtgtg gaatacatca caccctgcca cgcaggctgc tcaagctggg tggtccagga 1080 tgctctggac aacagccaga gtcctcccac ctcccaccct catgctgggc atcagcatct 1140 aaacctgagg ctcctccagg gagagacctg ggctgcactg gctggtgcag aagaacctgt 1200 tgatggtgca tagtccttca gaagccagcc aggcaccacc tgggcctgag agcccttcca 1260 gagaccccca ggccttggca ggtggagcag tgaactcctg tggatatggg aaccgattca 1320 aatccttctt aggcctctaa ctgactctgt taccttaggc aaattattta actagtgcct 1380 cagtttcttg gtctgtaaaa taggggagat attattaagt gcctactaca gagcaggaat 1440 gtgctgaata aatgctttac ctggatgaaa aaaaaaaaaa a 1481 <210> 31 <211> 667 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2656554CB1 <400> 31 ctaaagtggc agtgtttctt ctgaaattct caggcagtca gactgtctta ggcaaatctt 60 gataaaatag cccttatcca ggtttttatc taaggaatcc caagaagact ggggaatgga 120 gagacagtca agggttatgt cagaaaagga tgagtatcag tttcaacatc agggagcggt 180 ggagctgctt gtcttcaatt ttttgctcat ccttaccatt ttgacaatct ggttatttaa 240 aaatcatcga ttccgcttct tgcatgaaac tggaggagca atggtgtatg acaagccgcc 300 gaaatttgcc atgtcacgag agcaaatgtc acagtcatgt tctcacacgg cacataatgc 360 aagtctgttg acagatgcgg gtccattgtc atgtggggag tcgagggcga gctgtttgtt 420 tttgtaatga tgttgggaag tgatggctct gcagtcacaa agagcagcct tctctcactg 480 gctgcaccga tgaacattac gaagttctag aaaaaacatc acttcaaaat gcctggagta 540 attcctctta tatcaactaa tttcaagaag aaaacttgca gaaactaacc ccacccctct 600 taagagaata ttgtgtccaa gtccttttta tttatacgaa cagtgtctta ttttcttata 660 atgaaat <210> 32 <211> 1635 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2719228CB1 <400> 32 atagctgtct tgagccccaa gcctcttcct cccctgctgc ccctctgcag ccattcggga 60 tgggaccccc tctggggtgt cagcacgaaa gggctaacgg gagccccttc cttggcctcc 120 PCTNS99l26048 ccctgtaggt tacagagcca tcacggtgcg gaagctcatg cagggcatgg gccttggcct 180 ctccagcgtc tttgctctgt gcctgggcca cacctccagc ttctgtgagt ctgtggtctt 240 tgcatcagcc tccatcggcc tccagacctt caaccacagt ggcatttctg ttaa'catcca 300 ggacttggcc ccgtcctgcg ccggctttct gtttggtgtg gccaacacag ccggggcctt 360 ggcaggtgtc gtgggtgtgt gtctaggcgg ctacttgatg gagaccacgg gctcctggac 420 ttgcctgttc aaccttgtgg ccatcatcag caacctgggg ctgtgcacct tcctggtgtt 480 tggacaggct cagagggtgg acctgagctc tacccatgag gacctctagc tcccaacccc 540 acagcctctc caaggaccca ggcgccagca gccccgggac acaggggact cagtgtgtga 600 gacttggtca ctccatgtca gacacacgag cagagaggaa cacaaaccac tgtggagcct 660 gaagctcctt aagaagagtc cacaacagct ggtgggaggg tggggtgggc ctgggtccag 720 accaggctcg ctgctctctg ggcctcagtt tccccacctg ccagcgggct cggccctgtc 780 ctcctcacag gctggtgtgg ccgtcagggt gggtggggtt attgttagta ggcgcagcct 840 cattcccacc acgatctgtt ccgcgtggtt cccgccaaac ctccctcggt cgccgtgttc 900 tccgcaagcc tcctgcagcg cccgcctgcc aatgtgaggc tggcaccagg ctgcagcctc 960 cccaatccca gcccactttg ctgtgtctct ggcgggctgt cctccttggt gggagctgtc 1020 ctgcacactg taggatgctt aaaggtatcc ctggcctcca cccaccccta gccagcagct 1080 cccagtcaga caacagccag aaatgtctcc agactctgcc cagcctcccc aggtagccac 1140 cctcgagaca cgacctcaga gtctctgtgt ctcctagaag cctgacagag acccccaggg 1200 cagtgggtgg gtggcgggct agagaccctt gcctgtgtcc gggaccctgg cgccgctctc 1260 ccctcctgtg gatccctccg cactaacagt gttctcagtg ggcagacgcc tgggcacccc 1320 ttgggccctg cccagcatgg ccatggcgca ggctctcgaa cccgcatggc tttcccaggc 1380 ctggtgattc tgctctccag ggacggttgg caccttcctc gggggcgggc cccacgcacc 1440 ccagaacaca cagacccacc tttctggcgt tctttctacc tcccttttcg ttgcctgagg 1500 agctggtggt ttcatgagtt aatgatacat cttgcaaggt gtacacatag agaaaaaaac 1560 ctaaaaatgt ggaaaagcac gccaaagcct tatttaaata ataactatta aactattcaa 1620 aaagaaaaaa aaaaa <210> 33 <211> 1447 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 3657824CB1 <400> 33 ccgagcggtg ccaggccagg tgtgtgcgtc cgtcggtctt tccgtgccca cgccggagac 60 cagccccgga ggccgcctgg gcctatccct gtgccaggca ccatgaagca ggagtctgca 120 gccccgaaca ccccgcccac ctcgcagtcc cctacgccgt ccgctcagtt cccccgaaac 180 gacggcgacc ctcaagcgct gtggattttc gggtacggct ccctggtgtg gaggcccgac 240 ttcgcctaca gcgacagccg tgtgggcttc gtgcgcggct acagccgccg tttctggcag 300 ggagacacct tccatcgggg cagcgacaag atgcctggcc gtgtggtgac gctccttgaa 360 gatcatgagg gctgcacttg gggcgtggca taccaagtgc aaggggagca ggtaagcaag 420 gccctgaagt acctgaatgt gcgagaggca gtgcttggtg gctacgatac caaggaggtc 480 accttctatc cccaagatgc tcctgaccaa ccactgaagg cattggccta tgtggccacc 540 ccacagaacc ctggttacct gggccctgcg cctgaagagg ccattgccac gcagatcctg 600 gcctgccggg gcttctccgg ccacaacctt gaatacttgc tgcgtctggc agacttcatg 660 cagctctgtg ggcctcaggc gcaggacgag cacctggcag ccatcgtgga cgctgtgggc 720 accatgttgc cctgcttctg ccccaccgag caggctctgg cgctggtgtg aggggctgag 780 cccctgcggg gagtgctcat gtggacatca gggccagaca cccactccag tgcacaagac 840 agacttgcga ccgcttgagc ccactgagca gatatggtgg gtggctggag gcttctcttt 900 ctcagtccct gcctgtctgc cagcctgcag ctctcctgct tgacactgac ttactacttg 960 aaactttatt tattgcacca tgttggtgtg gtgggcaggt ggagggcctg ccctggacac 1020 aggggccctg ctgagcagtg gccccatcct ggaacttgac cagattcccc ccagtgctgc 1080 tgctaacccc acaccaccca ggcctccacc tccccaggga gtctccaaga gcctcgatcc 1140 tctgctcact cagcccagcc atccatagcc ctgggaattc cacctgccaa ggatoccagc 1200 aggctggatg agggatagta gggcatgagg agaaggagcc ctgtaaggac tgaggccccg 1260 gccagccctt ctcctccacc agttccccag agcagagctg gagctgatgc ctggacacag 1320 ctgctgagcc tggcctgggc ctcttaccca cttggttgtt ttcttgtccc tctgtctgtc 1380 tgtctatcta cttgtctgtc tgggccactc ctgcctgtgt gttggtctat tcctgggaag 1440 ctcatca <210> 34 <211> 657 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 5378485CB1 <400> 34 gactcctgtt gcgcatgctc agcgcgctgc ccggctgggg acccgcgcac ctgcagcgcc 60 cgctgctcgg ccctgcatcc tgcctgggca tcctgcgccc ggccatgacg gcgcactcat 120 tcgccctccc ggtcatcatc ttcaccacgt tctggggcct cgtcggcatc gccgggccct 180 ggttcgtgcc gaagggaccc aaccgcggag tgatcatcac catgctggtc gccaccgccg 240 tctgctgtta cctcttctgg ctcatcgcca tcctggcgca gctgaacccc ctgttcgggc 300 cccagctgaa gaatgagacc atctggtacg tgcgcttcct gtgggagtga cccgccgccc 360 ccgacccagg tgcccagctc tcggaatgac tgtggctcca ctgtccctga caaccccttc 420 gtccggaccc tcccccacac aactatgtct ggtcaccagc tccctcctgc tggcacccag 480 agacccggac ccgcaggccc tgcctggttc ctggaagtct tcccagtctt cccagccagc 540 ccggggccct ggggagccct gggcacagca gcggccgagg ggatgtcctg ctccaatact 600 cgcactgctc tggagtttgc actctttcgc aaggagatgc tgctggggag ctggtat 657 <210> 35 <211> 646 <212> PRT
<213> Mus musculus <300>
<308> GenBank ID No: 82612939 <400> 35 Met Arg Ala Pro Gly Ala Gly Thr Ala Ser Val Ala Ser Leu Ala Leu Leu Trp Phe Leu Gly Leu Pro Trp Thr Trp Ser Ala Ala Ala Ala Phe Cys Val Tyr Val Gly Gly Gly Gly Trp Arg Phe Leu Arg Ile Val Cys Lys Thr Ala Arg Arg Asp Leu Phe Gly Leu Ser Val Leu Ile Arg Val Arg Leu Glu Leu Arg Arg His Arg Arg Ala Gly Asp Thr Ile Pro Cys Ile Phe Gln Ala Val Ala Arg Arg Gln Pro Glu Arg Leu Ala Leu Val Asp Ala Ser Ser Gly Ile Cys Trp Thr Phe Ala Gln Leu Asp Thr Tyr Ser Asn Ala Val Ala Asn Leu Phe Arg Gln Leu Gly Phe Ala Pro Gly Asp Val Va1 Ala Val Phe Leu Glu Gly Arg Pro Glu Phe Val Gly Leu Trp Leu Gly Leu Ala Lys Ala Gly Val Val Ala Ala Leu Leu Asn Val Asn Leu Arg Arg Glu Pro Leu Ala Phe Cys Leu Gly Thr Ser Ala Ala Lys Ala Leu Ile Tyr Gly Gly Glu Met Ala Ala Ala Val Ala Glu Val Ser Glu Gln Leu Gly Lys Ser Leu Leu Lys Phe Cys Ser Gly Asp Leu Gly Pro Glu Ser Ile Leu Pro Asp Thr Gln Leu Leu Asp Pro Met Leu Ala Glu Ala Pro Thr Thr Pro Leu Ala Gln Ala Pro Gly Lys Gly Met Asp Asp Arg Leu Phe Tyr Ile Tyr Thr Ser Gly Thr Thr Gly Leu Pro Lys Ala Ala Ile Val Val His Ser Arg Tyr Tyr Arg Ile Ala Ala Phe Gly His His Ser Tyr Ser Met Arg Ala Ala Asp Val Leu Tyr Asp Cys Leu Pro Leu Tyr His Ser Ala Gly Asn Ile Met Gly Val Gly Gln Cys Val Ile Tyr Gly Leu Thr Val Val Leu Arg Lys Lys Phe Ser Ala Ser Arg Phe Trp Asp Asp Cys Val Lys Tyr Asn Cys Thr Val Val Gln Tyr Ile Gly Glu Ile Cys Arg Tyr Leu Leu Arg Gln Pro Val Arg Asp Val Glu Gln Arg His Arg Val Arg Leu Ala Val Gly Asn Gly Leu Arg Pro Ala Ile Trp Glu Glu Phe Thr Gln Arg Phe Gly Val Pro Gln Ile Gly Glu Phe Tyr Gly Ala Thr Glu Cys Asn Cys Ser Ile Ala Asn Met Asp Gly Lys Val Gly Ser Cys Gly Phe Asn Ser Arg Ile Leu Thr His Val Tyr Pro Ile Arg Leu Val Lys Val Asn Glu Asp Thr Met Glu Pro Leu Arg Asp Ser Glu Gly Leu Cys Ile Pro Cys Gln Pro Gly Glu Pro Gly Leu Leu Val Gly Gln Ile Asn Gln Gln Asp Pro Leu Arg Arg Phe Asp Gly Tyr Val Ser Asp Ser Ala Thr Asn Lys Lys Ile Ala His Ser Val Phe Arg Lys Gly Asp Ser Ala Tyr Leu Ser Gly Asp Val Leu Val Met Asp Glu Leu Gly Tyr Met Tyr Phe Arg Asp Arg Ser Gly Asp Thr Phe Arg Trp Arg Gly Glu Asn Val Ser Thr Thr Glu Val Glu pCT/US99l26048 Ala Val Leu Ser Arg Leu Leu Gly Gln Thr Asp Val Ala Va1 Tyr Gly Val Ala Val Pro Gly Val Glu Gly Lys Ala Gly Met Ala Ala Ile Ala Asp Pro His Ser Gln Leu Asp Pro Asn Ser Met Tyr Gln Glu Leu Gln Lys Val Leu Ala Ser Tyr Ala Arg Pro Ile Phe Leu Arg Leu Leu Pro Gln Val Asp Thr Thr Gly Thr Phe Lys Ile Gln Lys Thr Arg Leu Gln Arg Glu Gly Phe Asp Pro Arg Gln Thr Ser Asp Arg Leu Phe Phe Leu Asp Leu Lys Gln Gly Arg Tyr Val Pro Leu Asp Glu Arg Val His Ala Arg Ile Cys Ala Gly Asp Phe Ser Leu <210> 36 <211> 691 <212> PRT
<213> Schistosoma mansoni <300>
<308> GenBank ID No: g425474 <400> 36 Met Phe Ser Ala Leu Cys Arg Arg Gly Phe Leu Thr Asn Lys Val Ser Gln Phe Arg Ser Thr Tyr Lys Cys Asp His Tyr Asn Leu Lys Thr His Ile Lys Pro Leu Lys Cys Ser Ser Ser Leu Arg Leu Thr Val Gly Thr Gly Leu Phe Ile Ala Leu His Ser Lys Ile Ser Pro Glu Ser Arg Ile Gln Thr Val Gln Cys Glu Val Asp Ser Tyr Gln Thr Asp Gln Ile Thr Phe Ala Lys Ser Gly Gly Ile Pro Arg Tyr Ile Gly Val Leu Ile Leu Pro Asp Cys Val Tyr Leu Phe Gly Ala Ile Leu Gly Ala Phe Val Ala Ala Val Met Asn Val Tyr Ile Pro Leu Tyr Leu Gly Asp Phe Val Ser Ser Leu Ser Arg Cys Val Val Thr His Glu Gly Phe Val Ser Ala Val Tyr Val Pro Thr Leu Arg Leu Cys Ser Ser Tyr Leu Leu Gln Ser Leu Ser Thr Phe Leu Tyr Ile Gly Leu Leu Gly Ser Val Gly Glu Arg Met Ala Arg Arg Met Arg Ile Gln Leu Phe Arg Lys Leu Val Tyr Gln Asp Val Ala Tyr Phe Asp Val His Ser Ser Gly Lys Leu Val Glu Ile Ile Gly Ser Asp Val Gln Asn Phe Lys Ser Ser Phe Lys Gln Cys Ile Ser Gln Gly Leu Arg Asn Gly Ile Gln Val Val Gly Ser Val Phe Ala Leu Leu Ser Ile Ser Pro Thr Leu Thr Ala Ala Leu Ile Gly Cys Leu Pro Cys Val Phe Leu Ile Gly Ser Leu Met Gly Thr Glu Leu Arg His Ile Ser Arg Glu Val Gln Ser Gln Asn Ser Leu Phe Ala Ser Leu Ile Asp Glu Ala Phe Ser His Ile Arg Thr Val Lys Ser Leu Ala Met Glu Asp Phe Leu Ile Asn Lys Ile Asn Tyr Asn Val Asp Lys Ala Lys Met Leu Ser Glu Lys Leu Ser Phe Gly Ile Gly Ser Phe Gln Gly Leu Ser Asn Leu Thr Leu Asn Gly Val Val Leu Gly Val Leu Tyr Val Gly Gly His Leu Met Ser Arg Gly Glu Leu Asp Ala Gly His Leu Met Ser Phe Leu Ala Thr Thr Gln Thr Leu Gln 365 370 3?5 Arg Ser Leu Thr Gln Leu Ser Leu Leu Tyr Gly Gln Val Val Arg Gly Tyr Thr Ala Leu Lys Arg Ile His Asp Ile Leu Ala Leu Pro Ser Gly Ile Gly Ser Ile Pro Ser Ser Ser Ser Ser Leu Val Val Ser Lys Gln His Val Asn Asn Ile Lys Glu Leu Pro Ser Ser Ser Ile Tyr Ser Ala Pro Ser Ile Glu Phe Ser Asp Val Lys Phe Ala Tyr Pro Asn Arg Pro Glu Thr Ile Val Leu Asn Glu Leu Ser Met Phe Leu Pro Gly Gly Lys Val Ile Ala Leu Val Gly Gln Ser Gly 470 4?5 480 Ala Gly Lys Ser Thr Val Val Ser Leu Leu Glu Arg Phe Tyr Asp Pro Ile Ser Gly Glu Ile Leu Leu Asn Gly Asp Lys Leu Thr Asn Phe Asn Val Asn Tyr Leu Arg Ser Lys Leu Ile Gly Tyr Ile Ser Gln Glu Pro Gln Ile Phe Asn Ala Ser Ile Arg Glu Asn Ile Arg Phe Gly Arg Phe Asp Ala Thr Asp Glu Glu Val Glu Glu Ala Ala Lys Leu Ala Tyr Ala His Asp Phe Ile Ser Asn Asp Leu Pro Tyr Gly Tyr Asp Thr Leu Val Gly Gln Gly Thr Gly Thr Ile Ala Gly Leu Ser Gly Gly Gln Arg Gln Arg Ile Ala Ile Ala Arg Ile Leu Leu Lys Asri Ala Pro Ile Leu Leu Met Asp Glu Ala Thr Ser Ala Leu Asp Thr Glu Ser Glu Ala Lys Val Gln Asn Ala Leu Asn Asn_ A1a Met Lys Gly Arg Thr Val Leu Ile Ile Ala His Arg Leu Ser Thr Val Arg Lys Ala Asp Leu Ile Leu Val Met Ser Lys Gly Gln Ile Val Glu Lys Gly Thr His Ser Glu Leu Met Ala Asn His Gly Tyr Tyr Tyr Asn Leu Val Gln Arg Gln Glu Gly Cys Asp Val Phe Asp <210> 37 <211> 634 <212> PRT
<213> Rattus norvegicus <300>
<308> GenBank ID No: 83015617 <400> 37 Met Thr Val Ala Ser Thr Ala Ala Pro Ser Tyr Thr Thr Ser Asp Thr Asn Arg Val Ile Ser Thr Phe Ser Val Val Asp Tyr Val Val Phe Gly Leu Leu Leu Val Leu Ser Leu Val Ile Gly Leu Tyr His Ala Cys Arg Gly Trp Gly Arg His Thr Val Gly Glu Leu Leu Met Ala Asp Arg Lys Met Gly Cys Leu Pro Val Ala Leu Ser Leu Leu Ala Thr Phe Gln Ser Ala Val Ala Ile Leu Gly Gly Pro Ala Glu Ile Tyr Arg Phe Gly Thr Gln Tyr Trp Phe Leu Gly Cys Ser Tyr Phe Leu Gly Leu Leu Ile Pro Ala His Ile Phe Ile Pro Val Phe Tyr Arg Leu His Leu Thr Ser Ala Tyr Glu Tyr Leu Glu Leu Arg Phe Asn Lys Ala Val Arg Ile Cys Gly Thr Val Thr Phe Ile Phe Gln Met Val Val Tyr Met Gly Val Ala Leu Tyr Ala Pro Ser Leu Ala Leu Asn Ala Val Thr Gly Phe Asp Leu Trp Leu Ser Val Leu Ala Leu Gly Ile Val Cys Asn Ile Tyr Thr Ala Leu Gly Gly Leu Lys Ala Val Ile Trp Thr Asp Val Phe Gln Thr Leu Ile Met Phe Leu Gly Gln Leu Va1 Val Ile Ile Val Gly Ala Ala Lys Val Gly Gly Leu Gly His Val Trp Ala Val Ala Ser Gln His Gly Leu Ile pCT/US99/26048 Ser Gly Ile Glu Leu Asp Pro Asp Pro Phe Val Arg His Thr Phe Trp Thr Leu Ala Phe Gly Gly Val Phe Met Met Leu Ser Leu Tyr Gly Val Asn Gln Ala Gln Val Gln Arg Tyr Leu Ser Ser His Ser Glu Lys Ala Ala Val Leu Ser Cys Tyr Ala Val Phe Pro Cys Gln Gln Val Ala Leu Cys Met Sex Cys Leu Ile Gly Leu Val Met Phe Ala Tyr Tyr Lys Lys Tyr Ser Met Ser Pro Gln Gln Glu Gln Ala Ala Pro Asp Gln Leu Val Leu Tyr Phe Val Met Asp Leu Leu Lys Asp Met Pro Gly Leu Pro Gly Leu Phe Val Ala Cys Leu Phe Ser Gly Ser Leu Ser Thr Ile Ser Ser Ala Phe Asn Ser Leu Ala Thr Val Thr Met Glu Asp Leu Ile Gln Pro Trp Phe Pro Gln Leu Thr Glu Thr Arg Ala Ile Met Leu Ser Arg Ser Leu Ala Phe Ala Tyr Gly Leu Val Cys Leu Gly Met Ala Tyr Val Ser Ser His Leu Gly Ser Val Leu Gln Ala Ala Leu Ser Ile Phe Gly Met Val Gly Gly Pro Leu Leu Gly Leu Phe Cys Leu Gly Met Phe Phe Pro Cys Ala Asn Pro Leu Gly Ala Ile Val Gly Leu Leu Thr Gly Leu Thr Met Ala Phe Trp Ile Gly Ile Gly Ser Ile Val Ser Arg Met Ser Ser Ala Ala Ala Ser Pro Pro Leu Asn Gly Ser Ser Ser Phe Leu Pro Ser Asn Leu Thr Val Ala Thr Val Thr Thr Leu Met Pro Ser Thr Leu Ser Lys Pro Thr Gly Leu Gln Gln Phe Tyr Ser Leu Ser Tyr Leu Trp Tyr Ser Ala His Asn Ser Thr Thr Val Ile Ala Val Gly Leu Ile Val Ser Leu Leu Thr Gly Gly Met Arg Gly Arg Ser Leu Asn Pro Gly Thr Ile Tyr Pro Val Leu Pro Lys Leu Leu Ala Leu Leu Pro Leu Ser Cys Gln Lys Arg Leu Cys Trp Arg Ser His Asn Gln Asp Ile Pro Val Val Thr Asn Leu Phe Pro Glu Lys Met Gly Asn Gly Ala Leu Gln Asp Ser Arg Asp Lys Glu Arg Met Ala Glu Asp Gly Leu Val His Gln Pro Cys Ser Pro Thr Tyr Ile Val Gln Glu Thr Ser Leu <210> 38 <211> 507 <212> PRT
<213> Homo Sapiens <300>
<308> GenBank ID No: g3639058 <400> 38 Met Ala Gly Ala Gly Pro Lys Arg Arg Ala Leu Ala Ala Pro Ala Ala Glu Glu Lys Glu Glu Ala Arg Glu Lys Met Leu Ala Ala Lys Ser Ala Asp Gly Ser Ala Pro Ala Gly Glu Gly Glu Gly Val Thr Leu Gln Arg Asn Ile Thr Leu Leu Asn Gly Val Ala Ile Ile Val Gly Thr Ile Ile Gly Ser Gly Ile Phe Val Thr Pro Thr Gly Val Leu Lys Glu A1a Gly Ser Pro Gly Leu Ala Leu Val Val Trp Ala Ala Cys Gly Val Phe Ser Ile Val Gly Ala Leu Cys Tyr Ala Glu Leu Gly Thr Thr Ile Ser Lys Ser Gly Gly Asp Tyr Ala Tyr Met Leu Glu Val Tyr Gly Ser Leu Pro Ala Phe Leu Lys Leu Trp Ile Glu Leu Leu Ile Ile Arg Pro Sex Ser Gln Tyr Ile Val Ala Leu Val Phe Ala Thr Tyr Leu Leu Lys Pro Leu Phe Pro Thr Cys Pro Val Pro Glu Glu Ala Ala Lys Leu Val Ala Cys Leu Cys Val Leu Leu Leu Thr Ala Val Asn Cys Tyr Ser Val Lys Ala Ala Thr Arg Val Gln Asp Ala Phe Ala Ala Ala Lys Leu Leu Ala Leu Ala Leu Ile Ile Leu Leu Gly Phe Val Gln Ile Gly Lys Gly Asp Val Ser Asn Leu Asp Pro Lys Phe Ser Phe Glu Gly Thr Lys Leu Asp Val Gly Asn Ile Val Leu Ala Leu Tyr Ser Gly Leu Phe Ala Tyr Gly Gly TrP ~n Tyr Leu Asn Phe Val Thr Glu Glu Met Ile Asn Pro Tyr Arg Asn Leu Pro Leu Ala Ile Ile Ile Ser Leu Pro Ile Val Thr Leu Val Tyr Val Leu Thr Asn Leu Ala Tyr Phe Thr Thr Leu Ser Thr Glu Gln Met Leu Ser Ser Glu Ala Val Ala Val Asp Phe Gly Asn Tyr His Leu Gly Val Met Ser Trp Ile Ile Pro Val Phe Val Gly Leu Ser Cys Phe Gly Ser Val Asn Gly Ser Leu Phe Thr Ser Ser Arg Leu Phe Phe Val Gly Ser Arg Glu Gly His Leu Pro Ser Ile Leu Ser Met Ile His Pro Gln Leu Leu Thr Pro Val Pro Ser Leu Val Phe Thr Cys Val Met Thr Leu Leu Tyr Ala Phe Ser Lys Asp Ile Phe Ser Val Ile Asn Phe Phe Ser Phe Phe Asn Trp Leu Cys Val Ala Leu Ala Ile IIe Gly Met Ile Trp Leu Arg His Arg Lys Pro Glu Leu Glu Arg Pro Ile Lys Val Asn Leu Ala Leu Pro Val Phe Phe Ile Leu Ala Cys Leu Phe Leu Ile Ala Val Ser Phe Trp Lys Thr Pro Val Glu Cys Gly Ile Gly Phe Thr Ile Ile Leu Ser Gly Leu Pro Val Tyr Phe Phe Gly Val Trp Trp Lys Asn Lys Pro Lys Trp Leu Leu Gln Gly Ile Phe Ser Thr Thr Val Leu Cys Gln Lys Leu Met Gln Val Val Pro Gln Glu Thr <210> 39 <211> 504 <212> PRT
<213> Homo sapiens <300>
<308> GenBank ID No: 81840045 <400> 39 Met Glu Ala Pro Leu Gln Thr Glu Met Val Glu Leu Val Pro Asn Gly Lys His Ser Glu Gly Leu Leu Pro Val Ile Thr Pro Met Ala Gly Asn Gln Arg Val Glu Asp Pro Ala Arg Ser Cys Met Glu Gly Lys Ser Phe Leu Gln Lys Ser Pro Ser Lys Glu Pro His Phe Thr Asp Phe Glu Gly Lys Thr Ser Phe Gly Met Ser Val Phe Asn Leu Sex Asn Ala Ile Met Gly Ser Gly Ile Leu Gly Leu Ala Tyr Ala Met Ala Asn Thr Gly Ile Ile Leu Phe Leu Phe Leu Leu Thr Ala Val Ala Leu Leu Ser Ser Tyr Ser Ile His Leu Leu Leu Lys Ser Ser Gly Val Val Gly Ile Arg Ala Tyr Glu Gln Leu Gly Tyr Arg Ala Phe Gly Thr Pro Gly Lys Leu Ala Ala Ala Leu Ala Ile Thr pCT/US99/26048 Leu Gln Asn Ile Gly Ala Met Ser Ser Tyr Leu Tyr Ile Ile Lys 155 160 ~ 165 Ser Glu Leu Pro Leu Val Ile Gln Thr Phe Leu Asn Leu Glu Glu Lys Thr Ser Asp Trp Tyr Met Asn Gly Asn Tyr Leu Val Ile Leu Val Ser Val Thr Ile Ile Leu Pro Leu Ala Leu Met Arg Gln Leu Gly Tyr Leu Gly Tyr Ser Ser Gly Phe Ser Leu Ser Cys Met Val Phe Phe Leu Ile Ala Val Ile Tyr Lys Lys Phe His Val Pro Cys Pro Leu Pro Pro Asn Phe Asn Asn Thr Thr Gly Asn Phe Ser His Val GIu Ile Val Lys Glu Lys Val Gln Leu Gln Val Glu Pro Glu Ala Ser Ala Phe Cys Thr Pro Ser Tyr Phe Thr Leu Asn Ser Gln Thr Ala Tyr Thr Ile Pro Ile Met Ala Phe Ala Phe Val Cys His Pro Glu Val Leu Pro Ile Tyr Thr Glu Leu Lys Asp Pro Ser Lys Lys Lys Met Gln His Ile Ser Asn Leu Ser Ile Ala Val Met Tyr Ile Met Tyr Phe Leu Ala Ala Leu Phe Gly Tyr Leu Thr Phe Tyr Asn Gly Val Glu Ser Glu Leu Leu His Thr Tyr Ser Lys Val Asp Pro Phe Asp Val Leu Ile Leu Cys Val Arg Val Ala Val Leu Thr Ala Val Thr Leu Thr Val Pro Ile Val Leu Phe Pro Val Arg Arg Ala Ile Gln Gln Met Leu Phe Pro Asn Gln Glu Phe Ser Trp Leu Arg His Val Leu Ile Ala Val Gly Leu Leu Thr Cys Ile Asn Leu Leu Val Ile Phe Ala Pro Asn Ile Leu Gly Ile Phe GIy Val Ile Gly Ala Thr Ser Ala Pro Phe Leu Ile Phe Ile Phe Pro Ala Ile Phe Tyr Phe Arg Ile Met Pro Thr Glu Lys Glu Pro Ala Arg Ser Thr Pro Lys Ile Leu Ala Leu Cys Phe Ala Met Leu Gly Phe Leu Leu Met Thr Met Ser Leu Ser Phe Ile Ile Ile Asp Trp Ala Ser Gly Thr Ser Arg His Gly Gly Asn His <210> 40 <211> 393 <212> PRT
<213> Homo sapiens PC'fNS99126048 <300>
<308> GenBank ID No: g1526438 <400> 40 Met Ala Ala Val Gly Ala Gly Gly Ser Thr Ala Ala Pro Gly Pro Gly Ala Val Ser Ala Gly Ala Leu Glu Pro Gly Thr Ala Ser Ala Ala His Arg Arg Leu Lys Tyr Ile Ser Leu Ala Val Leu Val Val Gln Asn Ala Ser Leu Ile Leu Ser Ile Arg Tyr Ala Arg Thr Leu Pro Gly Asp Arg Phe Phe Ala Thr Thr Ala VaI Val Met Ala Glu Val Leu Lys Gly Leu Thr Cys Leu Leu Leu Leu Phe Ala Gln Lys Arg Gly Asn Val Lys His Leu Val Leu Phe Leu His Glu Ala Val Leu Val Gln Tyr Val Asp Thr Leu Lys Leu Ala Va1 Pro Ser Leu Ile Tyr Thr Leu Gln Asn Asn Leu Gln Tyr Val Ala Ile Ser Asn Leu Pro Ala Ala Thr Phe Gln Val Thr Tyr Gln Leu Lys Ile Leu Thr Thr Ala Leu Phe Ser Val Leu Met Leu Asn Arg Ser Leu Ser Arg Leu Gln Trp Ala Ser Leu Leu Leu Leu Phe Thr Gly Val Ala Ile Val Gln Ala Gln Gln Ala Gly Gly Gly Gly Pro Arg Pro Leu Asp Gln Asn Pro Gly Ala Gly Leu Ala Ala Val Val Ala Ser Cys Leu Ser Ser Gly Phe Ala Gly Val Tyr Phe Glu Lys Ile Leu Lys Gly Ser Ser Gly Ser Val Trp Leu Arg Asn Leu Gln Leu Gly Leu Phe Gly Thr Ala Leu Gly Leu Val Gly Leu Trp Trp Ala Glu Gly Thr Ala Val Ala Thr Arg Gly Phe Phe Phe Gly Tyr Thr Pro Ala Val Trp Gly Val Val Leu Asn Gln Ala Phe Gly Gly Leu Leu Val Ala Val Val Val Lys Tyr Ala Asp Asn Ile Leu Lys G1Y Phe Ala Thr Ser Leu Ser Ile Val Leu Ser Thr Val Ala Ser Ile Arg Leu Phe Gly Phe His Val Asp Pro Leu Phe Ala Leu Gly Ala Gly Leu Val Ile Gly Ala Val Tyr Leu Tyr Ser Leu Pro Arg Gly Ala Ala Lys Ala Ile Ala Sex Ala Ser Ala Ser Ala Ser Gly Pro Cys Val His Gln Gln Pro Pro Gly Gln Pro Pro Pro Pro Gln Leu Sex Ser His Arg Gly Asp Leu Ile Thr Glu Pro Phe Leu Pro Lys Ser Val Leu Val Lys ' <210> 41 <211> 893 <212> PRT
<213> Homo Sapiens <300>
<3pg> GenBank ID No: g3335175 <400> 41 His Val Gln Asp Phe Thr Ala Phe Trp Asp Lys Ala Ser Glu Thr Pro Thr Leu Gln Gly Leu Ser Phe Thr Val Arg Pro Gly Glu Leu Leu Ala Val Val Gly Pro Val Gly Ala Gly Lys Ser Ser Leu Leu Ser Ala Val Leu Gly Glu Leu Ala Pro Ser His Gly Leu Val Ser Val His Gly Arg Ile Ala Tyr Val Ser Gln Gln Pro Trp Val Phe 65 ?0 75 Ser Gly Thr Leu Arg Ser Asn Ile Leu Phe Gly Lys Lys Tyr Glu Lys Glu Arg Tyr Glu Lys Val Ile Lys Ala Cys Ala Leu Lys Lys Asp Leu Gln Leu Leu Glu Asp Gly Asp Leu Thr Val Ile Gly Asp Arg Gly Thr Thr Leu Ser Gly Gly Gln Lys Ala Arg Val Asn Leu Ala Arg Ala Val Tyr Gln Asp Ala Asp Ile Tyr Leu Leu Asp Asp Pro Leu Ser Ala Val Asp Ala Glu Val Ser Arg His Leu Phe Glu Leu Cys Ile Cys Gln Ile Leu His Glu Lys Ile Thr Ile Leu Val Thr His Gln Leu Gln Tyr Leu Lys Ala Ala Ser Gln Ile Leu Ile Leu Lys Asp Gly Lys Met Val Gln Lys Gly Thr Tyr Thr Glu Phe Leu Lys Ser Gly Ile Asp Phe Gly Ser Leu Leu Lys Lys Asp Asn Glu Glu Ser Glu Gln Pro Pro Val Pro Gly Thr Pro Thr Leu Arg Asn Arg Thr Phe Ser Glu Ser Ser Val Trp Ser Gln Gln Ser Ser Arg Pro Ser Leu Lys Asp Gly Ala Leu Glu Ser Gln Asp Thr Glu Asn Val Pro Val Thr Leu Ser Glu Glu Asn Arg Ser Glu Gly Lys Val Gly Phe Gln Ala Tyr Lys Asn Tyr Phe Arg Ala Gly Ala His Trp Ile Val Phe Ile Phe Leu Ile Leu Leu Asn Thr Ala Ala Gln Val Ala Tyr Val Leu Gln Asp Trp Trp Leu Ser Tyr Trp Ala Asn Lys Gln Ser Met Leu Asn Val Thr Val Asn Gly Gly Gly Asn Val Thr Glu Lys Leu Asp Leu Asn Trp Tyr Leu Gly Ile Tyr Ser Gly Leu Thr Val Ala Thr Val Leu Phe Gly Ile Ala Arg Ser Leu Leu Val Phe Tyr Val Leu Val Asn Ser Ser Gln Thr Leu His Asn Lys Met Phe GIu Ser Ile Leu Lys Ala Pro Val Leu Phe Phe Asp Arg Asn Pro Ile Gly Arg Ile Leu Asn Arg Phe Ser Lys Asp Ile Gly His Leu Asp Asp Leu Leu Pro Leu Thr Phe Leu Asp Phe Ile Glr1 Thr Leu Leu Gln Val Val Gly Val Val Ser Val Ala Val Ala Val Ile Pro Trp Ile Ala Ile Pro Leu Val Pro Leu Gly Ile Ile Phe Ile Phe Leu Arg Arg Tyr Phe Leu Glu Thr Ser Arg Asp Val Lys Arg Leu Glu Sex Thr Thr Arg Ser Pro Val Phe Ser His Leu Ser Ser Ser Leu Gln Gly Leu Trp Thr Ile Arg Ala Tyr Lys Ala Glu Glu Arg Cys Gln Glu Leu Phe Asp Ala His Gln Asp Leu His Ser Glu Ala Trp Phe Leu Phe Leu Thr Thr Ser Arg Trp Phe Ala Val Arg Leu Asp Ala Ile Cys Ala Met Phe Val Ile Ile Val Ala Phe Gly Ser Leu Ile Leu Ala Lys Thr Leu Asp Ala Gly Gln Val Gly Leu Ala Leu Ser Tyr Ala Leu Thr Leu Met Gly Met Phe Gln Trp Cys Val Arg Gln Ser Ala Glu Val Glu Asn Met Met Ile Ser Val Glu Arg Val Ile Glu Tyr Thr Asp Leu Glu Lys Glu Ala Pro Trp Glu Tyr Gln Lys Arg Pro Pro Pro Ala Trp Pro His Glu Gly Val Ile Ile Phe Asp Asn Val Asn Phe Met Tyr Ser Pro Gly Gly Pro Leu Val Leu Lys His Leu Thr Ala Leu Ile Lys Ser Gln Glu Lys Val Gly Ile Val Gly Arg Thr Gly Ala Gly Lys Ser Ser Leu Ile Ser Ala Leu Phe Arg Leu Ser Glu Pro Glu Gly Lys Ile Trp Ile Asp Lys Ile Leu Thr Thr Glu Ile Gly Leu His Asp Leu Arg Lys Lys Met Ser Ile Ile Pro Gln Glu Pro Val Leu Phe Thr Gly Thr Met Arg Lys Asn Leu Asp Pro Phe Lys Glu His Thr Asp Glu Glu 725 730 735, Leu Trp Asn Ala Leu Gln Glu Val Gln Leu Lys Glu Thr Ile Glu Asp Leu Pro Gly Lys Met Asp Thr Glu Leu Ala Glu Ser Gly Ser Asn Phe Ser Val Gly Gln Arg Gln Leu Val Cys Leu Ala Arg Ala Ile Leu Arg Lys Asn Gln Ile Leu Ile Ile Asp Glu Ala Thr Ala Asn Val Asp Pro Arg Thr Asp G1u Leu Ile Gln Lys Lys Ile Arg Glu Lys Phe Ala His Cys Thr Val Leu Thr Ile Ala His Arg Leu Asn Thr Ile Ile Asp Ser Asp Lys Ile Met Val Leu Asp Ser Gly Arg Leu Lys Glu Tyr Asp Glu Pro Tyr Val Leu Leu Gln Asn Lys Glu Ser Leu Phe Tyr Lys Met Val Gln Gln Leu Gly Lys Ala Glu 860 865 8?0 Ala Ala Ala Leu Thr Glu Thr Ala Lys Gln Val Ile Leu Gln Lys Lys Leu Ser Thr Tyr Trp Ser His <210> 42 <211> 453 <212> PRT
<213> Homo Sapiens <300>
<308> GenBank ID No: g1279457 <400> 42 Met Ala Leu Arg Gly Phe Cys Ser Arg Trp Leu Arg Pro Ala Leu Ala Ile Gly Leu Phe Ala Ser Met Ala Ala Val Leu Leu Gly Gly Ala Arg Ala Ser Arg Leu Leu Phe Gln Arg Leu Leu Trp Asp Val Val Arg Ser Pro Ile Ser Phe Phe Glu Arg Thr Pro Ile Gly His Leu Leu Asn Arg Phe Ser Lys Glu Thr Asp Thr Val Asp Val Asp Ile Pro Asp Lys Leu Arg Ser Leu Leu Met Tyr Ala Phe Gly Leu Leu Glu Val Ser Leu Val Val Glu Trp Pro Thr Pro Leu Pro Leu Trp Pro Ser Cys His Cys Phe Ser Ser Thr Leu Gly Phe Arg Trp Leu Ala Ala Asn Val Glu Leu Leu Gly Asn Gly Leu Val Phe Ala Ala Ala Thr Cys Ala Val Leu Ser Lys Ala His Leu Ser Ala Gly Leu Val Gly Phe Ser Val Ser Ala Ala Leu Gln Val Thr Gln Thrr Leu Gln Trp Val Val Arg Asn Trp Thr Asp Leu Glu Asn Ser Ile Val Ser Val Glu Arg Met Gln Asp Tyr Ala Trp Thr Pro Lys Glu Ala Pro Trp Arg Leu Pro Thr Cys Ala Ala Gln Pro Pro Trp Pro Gln Gly Gly Gln Ile Glu Phe Arg Asp Phe Gly Leu Arg Tyr Arg Pro Glu Leu Pro Leu Ala Val Gln Gly Val Ser Phe Lys Ile His Ala Gly Glu Lys Val Gly Ile Val Gly Arg Thr Gly Ala Gly Lys Ser Ser Leu Ala Ser Gly Leu Leu Arg Leu Gln Glu Ala Ala Glu Gly Gly Ile Trp Ile Asp Gly Val Pro Ile Ala His Val Gly Val His Thr Leu Arg Ser Arg Ile Ser Ile Ile Pro Gln Asp Pro Ile Leu Phe Pro Gly Ser Leu Arg Met Asn Leu Asp Leu Leu Gln Glu His Ser Asp Glu Ala Ile Trp Ala Ala Leu Glu Thr Val Gln Leu Lys Ala Leu Val Ala Cys Leu Pro Gly Gln Leu Gln Tyr Lys Cys Ala Asp Arg Gly Glu Asp Leu Ser Val Gly Gln Lys Gln Leu Leu Cys Leu Ala Arg Ala Leu Leu Arg Lys Thr Gln Ile Leu Ile Leu Asp Glu Ala Thr Ala Ala Val Asp Pro Gly Thr Glu Leu Gln Met Gln Ala Met Leu Gly Ser Trp Phe Ala Gln Cys Thr Val Leu Leu Ile Ala His Arg Leu Arg Ser Val Met Asp Cys Ala Arg Val Leu Val Met Asp Lys Gly Gln Val Ala Glu Ser Gly Ser Pro Ala Gln Leu Leu Ala Gln Lys Gly Leu Phe Tyr Arg Leu Ala Gln Glu Ser Gly Leu Val

Claims (20)

What is claimed is:

1. A substantially purified polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-17 and fragments thereof.

2. A substantially purified variant having at least 90% amino acid sequence identity to the amino acid sequence of claim 1.

3. An isolated and purified polynucleotide encoding the polypeptide of claim 1.

4. An isolated and purified polynucleotide variant having at least 90%
polynucleotide sequence identity to the polynucleotide of claim 3.

5. An isolated and purified polynucleotide which hybridizes under stringent conditions to the polynucleotide of claim 3.

6. An isolated and purified polynucleotide having a sequence which is complementary to the polynucleotide of claim 3.

7. A method for detecting a polynucleotide, the method comprising the steps of:
(a) hybridizing the polynucleotide of claim 6 to at least one nucleic acid in a sample, thereby forming a hybridization complex; and (b) detecting the hybridization complex, wherein the presence of the hybridization complex correlates with the presence of the polynucleotide in the sample.

8. The method of claim 7 further comprising amplifying the polynucleotide prior to hybridization.

9. An isolated and purified polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO:18-34 and fragments thereof.

10. An isolated and purified polynucleotide variant having at least 90%
polynucleotide sequence identity to the polynucleotide of claim 9.

11. An isolated and purified polynucleotide having a sequence which is complementary to the polynucleotide of claim 9.

12. An expression vector comprising at least a fragment of the polynucleotide of claim 3.

13. A host cell comprising the expression vector of claim 12.

14. A method for producing a polypeptide, the method comprising the steps of:
a) culturing the host cell of claim 13 under conditions suitable for the expression of the polypeptide; and b) recovering the polypeptide from the host cell culture.

15. A pharmaceutical composition comprising the polypeptide of claim 1 in conjunction with a suitable pharmaceutical carrier.

16. A purified antibody which specifically binds to the polypeptide of claim 1.

17. A purified agonist of the polypeptide of claim 1.

18. A purified antagonist of the polypeptide of claim 1.

19. A method for treating or preventing a disorder associated with decreased expression or activity of MTRP, the method comprising administering to a subject in need of such treatment an effective amount of the pharmaceutical composition of claim 15.

20. A method for treating or preventing a disorder associated with increased expression or activity of MTRP, the method comprising administering to a subject in need of such treatment an effective amount of the antagonist of claim 18.