JPH11308995A - New proteins c16n-1 and c16n-2, and genes coding for these - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain new genes which consists of DNA coding for proteins having specific amino acid sequences, have an activity of inducing differentiation from a myeloid cell to a cell having an ability of degrading hydroxyapatite and so on, and can be used, for example, for preparing an animal model for osteoporosis. SOLUTION: These are new DNA coding for new proteins C16N-1 and C16N-2 having amino acid sequences shown, for example, by formulas I and II which have an activity of inducing differentiation from a myeloid cell to a cell having an ability of degrading hydroxyapatite, an activity of keeping viability of nerve cells, an activity of suppressing growth of osteoblasts, an activity of enhancing expression of collagen type I in osteoblast, and so on, and are useful to prepare a transgenic animal useful as an animal model for developing medicines for hypertension and osteoporosis, and as an animal for screening the medicines and so on. This DNA is obtained by screening cDNA libraries from human brain, hippocampus, and testis using a part of DNA of C16N as a probe or a PCR primer.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to novel proteins C16N-1 and C16N-2, or genes encoding them. More specifically, unlike C16N, non-secretory proteins C16N-1 and C16N-2, or their analogous proteins, genes encoding these proteins, expression vectors of the genes, and traits transformed by the expression vectors The present invention relates to a transformant, a method for producing a recombinant protein using the transformant, a drug containing the gene or the protein as an active ingredient, an antibody against the protein, and a transgenic animal related to the introduction and deletion of the gene.

[0002]

2. Description of the Related Art The present inventors have been searching for an osteoclast differentiation-inducing factor from BW5147 cells, which are bone metastatic cells, by the expression cloning method. In the process, it is different from the osteoclast differentiation-inducing factor, but has a novel activity of inducing the differentiation of bone marrow cells into cells that degrade hydroxyapatite (crystals of bone calcium) and release calcium into the blood. The protein, C16N, was cloned (WO 97/40150). As a result of the analysis, the C16N gene was specifically expressed in bone and brain, and in addition to the above-mentioned activities, the activity of maintaining survival of nerve cells, the activity of inhibiting the growth of osteoblasts, and the type I collagen in osteoblasts It has also been found that it also has an expression promoting activity. Therefore, it is considered that C16N acts in vivo as a regulator of blood calcium concentration, as a neurotrophic factor, and as a promoter of osteoblast differentiation or function (International Patent Publication WO 97/97). / 40150). C16N has also been shown to be a secreted protein (WO97
/ 40150). However, besides this C16N, C16N
It has not yet been reported that a non-secreted protein exists as a structurally similar factor.

[0003]

An object of the present invention is to provide C16N-1 and C16N-2, non-secreted C16N-like factors that have not been cloned, or genes encoding them. That is, the present invention provides novel proteins C16N-1 and C16N-1.
N-2 or its analogous proteins, genes encoding these proteins, expression vectors of the genes, transformants transformed by the expression vectors, methods for producing recombinant proteins using the transformants, the genes Alternatively, an object of the present invention is to provide a medicine containing the protein as an active ingredient, an antibody against the protein, and a transgenic animal related to the introduction and deletion of the gene.

[0004]

Means for Solving the Problems The present inventors have examined the possibility that a structural similar factor of C16N exists in a living body as part of an analysis study of C16N. That is, C
Using a portion of the 16N DNA as a PCR primer, 5 ′
The C16N by the RACE method and the 3′-RACE method
A search for similar factors was performed. As a result, we succeeded in obtaining a DNA encoding the novel C16N-like protein having the entire amino acid sequence of C16N and further having the N-terminal side 150 amino acids or 133 amino acids longer than C16N. We named this novel C16N-like protein a protein whose N-terminal side is 150 amino acids longer, C16N-1,
Those whose N-terminal side was 133 amino acids longer were named C16N-2.

[0005] The N-terminal portion of the C16N-1 and C16N-2 of the present invention, ie, the novel 150 amino acid (C16N-1) or 133 amino acid (C16N-2) at the N-terminal, which is not present in C16N. As a result of performing a hydrophobicity plot on the portion, it was revealed that a region having a high hydrophobicity was present in these N-terminal side portions. Therefore, C16N-1 and C16N-2 of the present invention were present on the cell membrane. It was expected that the protein would be Also,
As a result of Western blot analysis using crushed mouse brain tissue and osteoblast cells, the C16N-
1 and C16N-2 were not secreted proteins, but were expected to be proteins present in cells or in cell membranes. From the above results, C16N-1 and C16N-1 of the present invention
16N-2, unlike C16N, which is a secreted protein, is thought to be a non-secreted protein and probably a membrane protein. The present invention has been completed based on the above findings.

That is, the gist of the present invention is: 1) a DNA encoding a protein consisting of the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6 or SEQ ID NO: 8, 2) SEQ ID NO: 1 , SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 7;
3) A DNA which hybridizes with the DNA according to 1) or 2) under stringent conditions and has the following properties (1), (2), (3) and / or (4): ) Encoding a non-secreted protein having
A, (1) having an activity of inducing differentiation of bone marrow cells into cells having hydroxyapatite decomposability, (2) having an activity of maintaining survival of nerve cells, (3) having an activity of inhibiting osteoblast proliferation, (4) Has the activity of promoting the expression of type I collagen in osteoblasts. 4) deletion of one or more amino acids in the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6 or SEQ ID NO: 8 , Consisting of an amino acid sequence substituted and / or added, and having the following properties (1), (2),
DNA encoding a non-secretory protein having (3) and / or (4), (1) having an activity of inducing differentiation from bone marrow cells to cells having hydroxyapatite decomposability, and (2) maintaining neuronal cell survival. (3) having osteoblast proliferation inhibitory activity; (4) having type I collagen expression promoting activity in osteoblasts; and 5) expressing the DNA according to any of 1) to 4) above. 6) an expression vector containing the DNA of any one of 1) to 4) above; 7) a transformant transformed by the expression vector of 6) above; 8) a trait of 7) above A method for producing a recombinant protein, comprising culturing the transformant and collecting the expressed recombinant protein. 9) The DNA according to any one of 1) to 4) above. Or a drug containing the protein of the above 5) as an active ingredient; 10) an antibody against the protein of the above 5); and 11) the DNA of any of the above 1) to 4) artificially introduced into a chromosome. Or a transgenic animal in which either is deleted from the chromosome.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, DNA is C1
6N-1 or C16N-2 DNA or the C16
DNAs similar to the DNAs of N-1 and C16N-2, and (1) an activity of inducing differentiation from bone marrow cells to cells having hydroxyapatite decomposability, (2) an activity of maintaining survival of nerve cells, (3) a bone The DNA is not particularly limited as long as it has a blast proliferation inhibitory activity and / or (4) a type I collagen expression promoting activity in osteoblasts and encodes a non-secretory protein.

[0008] As specific examples of the DNA of the present invention, for example, a DNA encoding a protein consisting of the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6 or SEQ ID NO: 8, or SEQ ID NO: 1 , SEQ ID NO: 3, SEQ ID NO: 5 or SEQ ID NO: 7 (hereinafter, these DNAs may be referred to as mouse and human C16N-1 and C16N-2 DNAs). Can be Furthermore, DNAs that hybridize under stringent conditions to any of these mouse and human C16N-1 and C16N-2 DNAs, or mouse and human C16N-1 and C16N-2 DNAs
DNA encoding a protein consisting of an amino acid sequence in which one or more amino acids have been deleted, substituted and / or added to any of the amino acid sequences described in (1) above, A non-secretory type having differentiation-inducing activity, (2) activity for maintaining survival of nerve cells, (3) activity for suppressing osteoblast proliferation, and / or (4) activity for promoting the expression of type I collagen in osteoblasts The DNA of the present invention is included as long as it is a DNA that encodes the above-mentioned protein. Hereinafter, these DNAs of the present invention will be sequentially described.

1) Mouse and human forms of C16N-1 and
DNA encoding C16N-2 Among the DNAs of the present invention, "DNA encoding a protein consisting of the amino acid sequence of SEQ ID NO: 2, or DNA consisting of the base sequence of SEQ ID NO: 1" DNA encoding mouse C16N-1 of the present invention
It is. Further, “DNA encoding a protein consisting of the amino acid sequence of SEQ ID NO: 4, or SEQ ID NO: 3
The DNA having the nucleotide sequence described in (1) is a DNA encoding the mouse C16N-2 of the present invention. The “DNA encoding the protein having the amino acid sequence of SEQ ID NO: 6 or the DNA having the nucleotide sequence of SEQ ID NO: 5” is a DNA encoding the human C16N-1 of the present invention. . "A DNA encoding a protein consisting of the amino acid sequence of SEQ ID NO: 8,
Alternatively, "a DNA comprising the nucleotide sequence of SEQ ID NO: 7" is a DNA encoding the human C16N-2 of the present invention.

DNAs encoding these mouse and human C16N-1 and C16N-2 are described, for example, in WO97.
For example, by using a part of the C16N DNA described in / 40150 as a probe or a PCR primer, a cDNA library of mouse or human brain, hippocampus, testis or the like can be screened for cloning. The cloning is performed, for example, by Molecul
ar Cloning 2nd Edt.Cold Spring Harbor Laboratory P
According to ress (1989) and the like, those skilled in the art can easily carry out the method. More specifically, the 5'-RACE method (Marathon-Rea
By performing dy cDNA Amprification Kit (manufactured by CLONTECH)), a DNA encoding the N-terminal 150 amino acid portion (C16N-1) or 133 amino acid portion (C16N-2) which is not present in C16N can be obtained. . This newly obtained DNA portion was replaced with a DNA portion encoding C16N (the DNA portion previously obtained by the 5'-RACE method).
Part and the following part), C16N-1
And C16N-2-encoding DNA.

2) mouse and human forms of C16N-1 and
Any of the DNAs encoding C16N-2 and a string
DNA that hybridizes under gentle conditions Among the DNAs of the present invention, “mouse and human C1
DNA that hybridizes with 6N-1 and C16N-2 DNA under stringent conditions, and (1) activity of inducing differentiation from bone marrow cells to cells having hydroxyapatite decomposability, (2) activity of maintaining survival of nerve cells (3) D which has an activity of inhibiting the growth of type I collagen in osteoblasts and / or (4) an activity of promoting the expression of type I collagen in osteoblasts, and / or which encodes a non-secretory protein
The term "NA" includes C16N-1 DNA and C16N-2 DNA of all vertebrates other than mice and humans.

Here, “DNA that hybridizes under stringent conditions” refers to, for example, standard hybridization conditions (formamide concentration: 50%,
(Salt concentration: 5 × SSC, temperature: 42 ° C.), the D of mouse or human C16N-1 and C16N-2
Refers to DNA that hybridizes with NA. These DNAs are, for example, mouse or human C16N-
1. Cloning can be performed by hybridization using a part of the C16N-2 DNA as a probe, or by a PCR method using a part of the DNA as a primer. Specific procedures for preparing a cDNA library, hybridization, selection of positive colonies, determination of a base sequence, and the like are all known, for example,
lecular Cloning 2nd Edt.Cold Spring Harbor Labora
This can be easily performed with reference to tory Press (1989) and the like. Examples of the cDNA library include various vertebrate brain, hippocampus, or testis-derived cDNA libraries.

Here, "activity of inducing differentiation of bone marrow cells into cells having hydroxyapatite degradability", "activity of maintaining survival of nerve cells", "activity of inhibiting proliferation of osteoblasts", "activity of type I collagen in osteoblasts" `` Expression promoting activity ''
All can be measured by the individual activity measurement methods described in WO97 / 40150. However, since the protein of the present invention is a non-secreted protein, and is probably considered to be a membrane protein, a membrane fraction of cells into which the DNA of the present invention has been introduced is prepared as described below, and this membrane fraction is The above activity can be measured by adding a sample solubilized with a surfactant or the like to a cell for assay as a sample. Furthermore, instead of using a protein as a sample, the above activity can also be measured by directly transfecting the DNA of the present invention into cells for assay. Here, the gene can be introduced into the cells for assay, for example, by the DEAE-dextram method, the calcium phosphate method, or the like.

[0014] Whether or not the protein is a "non-secretory protein" can be determined by, for example, the following method. That is, the measurement can be performed by fixing the cells on a slide chamber and examining the localization in the cells by reaction with a fluorescently labeled anti-C16N antibody. Alternatively, it can be measured by analyzing cells stained with a fluorescent antibody by flow cytometry.

3) mouse and human C16N-1 and
Encodes any modified protein of C16N-2
DNA Among the above DNAs, “mouse and human C16N-1
And one or more amino acids of any of the amino acid sequences of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8 (SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 8). And / or (1) an activity of inducing differentiation of bone marrow cells into cells having the ability to degrade hydroxyapatite, (2) an activity of maintaining the survival of nerve cells, (3) an activity of inhibiting the growth of osteoblasts, And / or (4) a DNA which has the activity of promoting the expression of type I collagen in osteoblasts and encodes a non-secretory protein " DNA encoding a non-secretory protein having the above activity among allelic variants and the like
Point to. DNA encoding the protein can be obtained, for example, by site-directed mutagenesis (Methods in Enzymology, 100, p468).
(1983)) and PCR (Molecular Cloning 2nd Edt.
Chapters, Cold Spring Harbor Laboratory Press (1989)), etc., can be easily prepared by those skilled in the art. Here, the number of amino acid residues to be deleted, substituted and / or added refers to the number of amino acids that can be deleted, substituted and / or added by a well-known method such as site-directed mutagenesis.

In the present invention, the protein is a protein obtained by expressing the various DNAs of the present invention. As a specific example, for example, SEQ ID NO:
Mouse C16N-1 having the amino acid sequence of 2,
Mouse C1 having the amino acid sequence of SEQ ID NO: 4
6N-2, human C16N-1 having the amino acid sequence of SEQ ID NO: 6, or human C16N-2 having the amino acid sequence of SEQ ID NO: 8.

These proteins are, for example, the D
It can be obtained by ligating NA to a known expression vector such as pBK-CMV, and then introducing it into an appropriate host for expression and production. The host may be either a prokaryotic cell or a eukaryotic cell. For example, Escherichia coli strains and animal cell strains are already widely spread and readily available unless otherwise specified. For example, animal cell hosts include COS-1, COS-7, and CHO cells. To transform an appropriate animal cell host using the expression plasmid, the LIPOFECTIN method (Felgner PL, et al, Pro
c. Natl. Acad. Sci. USA, 84, p7413 (1987)). It is likely that the protein of the present invention is present in the cell membrane fraction of the transformed cells, and the cell membrane fraction contains the protein of the present invention to such an extent that it can be used for various assays. Therefore, for example, fractionation of the intracellular membrane structure by centrifugation (Yoshio Imai et al., Cell fractionation method, Shinsei Kagaku Jikkensho Koza Lecture 1, Protein I, separation, purification, and properties (Tokyo Kagaku Dojin) p40-52 (1990))
Using the cell membrane fraction prepared by the above method, various activities of the protein of the present invention can be measured.

The DNA and protein of the present invention also include
It can be used as an active ingredient of a medicine. Specifically, diseases accompanied by a decrease in blood calcium concentration, such as hypocalcemia described in WO97 / 40150, massive blood loss due to massive bleeding and radiation exposure, or Parkinson's disease and Alzheimer's disease Neurological disorders, such as
Alternatively, it can be used for bone diseases such as osteoporosis, spinal cord injury, and fracture.

In particular, since the protein of the present invention is a non-secretory protein, it has an advantage that its action can be limited to a specific tissue or a specific site, as compared with C16N which is a secretory protein. Therefore, for example, in the case of a severe fracture or a complicated fracture, a repair treatment by an ex vivo method is performed after removing own bone (osteoblasts and chondrocytes), introducing the DNA of the present invention, and returning the cells to a defective portion. In such a case, it is considered that the action of the protein of the present invention can be limited around the introduced cells, and thus it is possible that the fracture can be repaired more efficiently than C16N.

As a method for introducing such a gene therapy agent containing the DNA of the present invention as an active ingredient into a cell, a gene transfer method using a viral vector or a non-viral gene transfer method (Experimental Medical Supplement) Any of the methods described in “Basic Technology for Gene Therapy”, Youtosha (1996) can be applied. In the above-mentioned repair treatment, it is desirable that the vector disappears or is inactivated immediately after the completion of the repair. Therefore, the transient introduction method is more preferable. Examples of a method for introducing a gene using a viral vector include a method of incorporating the DNA of the present invention into an adenovirus, a herpes virus, a vaccinia virus, a pox virus, a polio virus, a simbis virus, or another DNA virus, or an RNA virus, and introducing the DNA. Can be Among them, a method using adenovirus or vaccinia virus is particularly preferred. Examples of the non-viral gene transfer method include a liposome method, a lipofectin method, a microinjection method, a calcium phosphate method, and an electroporation method, and the liposome method is particularly preferable.

In the present invention, an antibody is an antibody against the protein of the present invention. The antibody can be easily prepared, for example, by immunizing a rabbit or the like using the method described in New Cell Engineering Experiment Protocol p210 Shujunsha (1993). In addition, for example, a monoclonal antibody can be easily produced by using the method described in Protein Experimental Methods for Molecular Biology Research, Chapter 4, Yodosha (1994). Furthermore, a humanized antibody can be used according to JP-A-62-296890. Uses of the antibody include affinity chromatography, screening of a cDNA library, diagnostic reagents and experimental reagents.

In the present invention, the term "transgenic animal" refers to a so-called transgenic animal in which the DNA of the present invention is artificially introduced into a chromosome, or a so-called knockout animal in which a chromosome is deleted. These transgenic animals are, for example, disease model mice: Molecu
A person skilled in the art can easily produce it based on lar Medicine special issue Nakayama Shoten (1994) and the like. The transgenic animal is very useful as a model animal for drug development such as hypertension and osteoporosis, or as an animal for screening the drug.

[0023]

EXAMPLES Hereinafter, the present invention will be described in more detail by way of examples as examples of the present invention, but the present invention is not limited to these examples.

Example 1 Residues of mouse and human C16N-1 and C16N-2
The following experiments were performed in order to examine the presence of genes encoding C16N analogous proteins of mouse and human clones. That is, in order to examine the presence of a gene encoding a mouse C16N-like protein, WO97 / 4
Mouse C16 described in SEQ ID NO: 3 in the sequence listing of No. 0150
The antisense DNA (R24) (5'-CGAGAGCCAGCCGTACCTCC-3 ') at positions 732 to 752 of the N base sequence, the antisense DNA (R9) at positions 686 to 705, and the positions 1779 to 1779 The sense DNA at position 1798 (F
7) (5'-AGGAGCTGCTACAACTGCTA-3 ') and 2078th ~
2097th position sense DNA (F14) (5-AACACCCCAG
CTTTCTGGCT-3) as PCR primer and Marathon-Rea
dy cDNA Amplification Kit [mouse brain, ovary, testis,
Embryo] (1st PC with AP1 and R24 primer set as 5 'RACE method according to the protocol of CLONTECH)
R, and using the PCR product as a template, AP
A second PCR was performed with the 2 and R9 primer sets.
Next, as a 3 ′ RACE method, the first PCR was performed with the F7 and AP2 primer sets, and the PCR product was used as a template, and the second PCR was performed with the F14 and AP2 primer sets.
CR was performed. From the above experiments, it was examined whether a cDNA clone longer in the 5 ′ direction or 3 ′ direction than C16N was obtained. As a result, a cDNA clone longer in the 3 ′ direction was not obtained, but two clones longer in the 5 ′ direction than C16N were obtained. These two types of clones were each used in a TA cloning vector (pGEM-T Easy
Vector) and confirm the nucleotide sequence.
Cleavage with restriction enzymes SalI and BglII, mouse C16N
Plasmid (SEQ ID NO: 3) was ligated to a plasmid cleaved with the same restriction enzyme, and pBK / mC1 containing a cDNA for a novel mouse protein C16N-1
6BK-1 and pBK / mC16N-2 having cDNA of a novel mouse protein C16N-2 were obtained.

Human C16N-1 and C16N-
DNA No. 2 was obtained as follows. That is,
Since the human and mouse C16N nucleotide sequences have extremely high homology, they can be used even when the primers used for PCR of mouse DNA are human. Therefore, human C1
In order to examine the presence of the gene encoding the 6N analogous protein, SEQ ID NO:
3. The antisense DNA (R24) (5′-CGAGAGC) at positions 732 to 752 of the nucleotide sequence of mouse C16N described in Item 3
CAGCCGTACCTCC-3 ') and the antisense DNA (R9) at positions 686 to 705, and positions 1779 to 1798
Position sense DNA (F7) (5'-AGGAGCTGCTACAACTGCTA-
3 ') and the nucleotide sequence of mouse C16N at position 2078 to position 2 of SEQ ID NO: 3 in WO97 / 40150.
097 position sense DNA (F14) (5-AACACCCCAGCTTT
CTGGCT-3) as PCR primer and Marathon-Ready c
DNA Amplification Kit [Human brain, hippocampus, ovary, testis]
According to the protocol of CLONTECH, the first PCR was performed with the AP1 and R24 primer sets as a 5 ′ RACE method, and the second PCR was performed with the AP2 and R9 primer sets using the PCR product as a template. Next, the first PCR was performed using the F7 and AP2 primer sets as a 3 ′ RACE method, and the PCR product was used as a template to perform the second PCR using the F14 and AP1 primer sets.
Was done. From the above experiments, it was examined whether a cDNA clone longer in the 5 ′ direction or 3 ′ direction than C16N was obtained. As a result, a cDNA clone longer in the 3 ′ direction was not obtained, but two clones longer in the 5 ′ direction than C16N were obtained. These two types of clones were each subcloned into a TA cloning vector (pGEM-T Easy vector), and after confirming the nucleotide sequence, cleaved with restriction enzymes SalI and BglII to obtain human C16N cDNA.
A plasmid having NA (SEQ ID NO: 5) was ligated with a plasmid cleaved with the same restriction enzyme, and a novel human protein C
PBK / hC16N-1 having 16N-1 cDNA
Thus, pBK / hC16N-2 having the cDNA of the novel human protein C16N-2 was obtained.

Example 2 Salts of mouse and human C16N-1 and C16N-2
Determination of Base Sequences The base sequences of mouse and human C16N-1 and C16N-2 were determined using the AutoRead Sequencing kit (Pharmacia).
Biotech) according to the instructions attached to the kit,
(ALF redDNA automatic sequencer (Pharmacia Bi
otech). The plasmid pBK / m
C16N-1, pBK / mC16N-2, pBK / hC
Inserted cDNA of 16N-1 and pBK / hC16N-2
Was determined.

The nucleotide sequence of mouse C16N-1 is shown in SEQ ID NO: 1, its amino acid sequence is shown in SEQ ID NO: 2, the nucleotide sequence of mouse C16N-2 is shown in SEQ ID NO: 3, and its amino acid sequence is shown in SEQ ID NO: : 4. Furthermore, the nucleotide sequence of human C16N-1 is shown in SEQ ID NO: 5, its amino acid sequence is shown in SEQ ID NO: 6, the nucleotide sequence of human C16N-2 is shown in SEQ ID NO: 7, and its amino acid sequence is shown in SEQ ID NO: No. 8 is described. Mouse type C16N-1 and C16N
-2 is SEQ ID NO: 4 in the sequence listing of WO97 / 40150.
The amino acid level was 150 amino acids and 133 amino acids longer at the N-terminal side, respectively, as compared with mouse C16N described in (1). In addition, human C16N-1 and C16N-1
N-2 is also SEQ ID NO: in the sequence listing of WO97 / 40150.
As compared with the human C16N described in No. 6, the N-terminal side was 150 amino acids and 133 amino acids longer at the amino acid level, respectively.

Of these C16N-1 and C16N-2,
The novel 150 amino acids and 133 amino acids at the N-terminus were subjected to hydrophobicity plotting by the method of Kyaito-Little.
Mouse type C1 described in SEQ ID NO: 4 at the positions 53 to 64 and 102 to 117 of the amino acid sequence of -1
Positions 36 to 47 and 85 to 100 of the amino acid sequence of 6N-2, and positions 53 to 64 and 102 of human C16N-1 described in SEQ ID NO: 6 Human C16N as set forth in SEQ ID NO: 8 to a portion at position 117.
It was found that the 36th to 47th positions and the 85th to 100th positions of -2 were highly hydrophobic regions. Therefore, these proteins were expected to be membrane proteins.

Example 3 Genes Corresponding to C16N-1 and C16N-2 by PCR Analysis
Marat made from RNA prepared from the child expression various tissues
hon Ready cDNA (manufactured by CLONTECH)
Was used as a template to perform a nested PCR reaction. As a primer sequence for amplification of the present gene, AP1 (attached to cDNA) as a 5 ′ primer for the first PCR, and 5′-GCATCTTC as a 3 ′ primer
Using GGCTCTTTGGAAATCC-3 ′, a PCR reaction was performed according to a standard method. 1 / of the amplified PCR product
Using the amount of 10 as a template, the second PCR was performed using AP2 (attached to cDNA) as a 5 'primer and 5'-CTGGTCAAAAGCCGTAGCCG as a 3' primer.
Performed using TAGCC-3 '. 1/1 of the reaction mixture
A 0% 1% gel was electrophoresed on a Loose gel to confirm the band.

As a result, the gene corresponding to C16N-1 was expressed in the testis and embryo of the mouse. The gene corresponding to C16N-2 was expressed in all tissues examined, but specifically expressed in human and mouse brain.

Example 4 C16N-1 Protein by Western Blotting Analysis
Or examination of C16N-2 protein expression ATDC5 cells derived from mouse embryo, UMR derived from rat osteosarcoma
106 cells, mouse skull-derived primary cultured osteoblasts, rat costal cartilage-derived primary cultured chondrocytes, and mouse thymoma-derived BW5147 cells were cultured, and the culture supernatant (10-fold concentrated) and cell extracts, and mouse brain Extract 30μg each
Was separated by SDS-polyacrylamide gel electrophoresis, transferred to a membrane filter by electroblotting, reacted with an anti-C16N antibody, and then reacted with C16N according to a standard method.
The protein band was developed.

As a result, the extract of mouse brain tissue and ATDC5 cells contained C16N-1 or C16N-1 having a molecular weight of 80 kDa.
There was a protein corresponding to 16N-2. The culture supernatant of ATDC5 cells, BW5147 cells, primary cultured osteoblasts and chondrocytes contained a protein corresponding to conventional C16N having a molecular weight of 63 kDa. As described above, the molecular weight of the secretory C16N protein detected in the culture supernatant is 63 kDa, and the C16N protein present in the tissue or cell extract is
The molecular weight of the 16N-1 or C16N-2 protein was 80 kDa. C16 equivalent to this 80 kDa
The N-1 and C16N-2 proteins are considered non-secreted.

[0033]

According to the present invention, a novel protein C16
N-1 and C16N-2, genes encoding these proteins and the like are provided. Since these proteins are non-secretory proteins, they have the advantage that their effects can be limited to specific tissues and specific sites.

[0034]

[Sequence List] SEQ ID NO: 1 Sequence length: 3675 Sequence type: nucleic acid Number of strands: double-stranded Topology: linear Sequence type: cDNA Sequence characteristics Characteristics determined method: E sequence CGCGGCGTGA GCAGCAGCCC CAGGCTCCCG GAGCATCGCG CCCGGAGAAA GACTTCGCCG 60 CTCGGGGCCG CAGCCTGGTG AGCTCAGCCC CCTTCAAGCC CTCCCCTGCA TCTCAGCCGG 120 AGCCTCTCCG AACCGGCGCT GATCGATGCC GAGACTCCCC AGGGACCCTA TCGCGACTCC 180 ATCGTGCCAT ATCTCGACAT CACCGTACCC TGTCGAGACT CCATTTTGTC ACAACCCCTT 240 TCAATATTTA TCTATTATAT ATATTTTTAA AATTTGCCCT ATCATATTTG GGGGCTGTCC 300 CCTTCATGTC GTGATTTCGC TGTGATCTCT CCGTGACATC ACCGCGCCAT CGTGAAGTGT 360 GATCTCATCG CTGCCCTGTC GTTCGACTTC ATCA ATG TCG TGT TGT GAC CTG 412 Met Ser Cys Cys Asp Leu 5 GCT GCG GCG GGA CAG TTG GGC AAG GCG GGC ATC ATG GCC TCG GAT TGT 460 Ala Ala Ala Gly Gln Leu Gly Lys Ala Gly Ile Met Ala Ser Asp Cys 10 15 20 GAG CCA GCT CTG AAC CAG GCA GAG AGC CGA AAC CCC ACC CTG GAG CGC 508 Glu Pro Ala Leu Asn Gln Ala Glu Ser Arg Asn Pro Thr Le u Glu Arg 25 30 35 TAC CTG GGA GCC CTC CGT GAG GCC AAG AAT GAC AGC GAG CAG TTT GCA 556 Tyr Leu Gly Ala Leu Arg Glu Ala Lys Asn Asp Ser Glu Gln Phe Ala 40 45 50 GCC CTG CTG CTA GTA ACC AAG GCA GTC AAA GCA GGT GAC ATT GAC GCC 604 Ala Leu Leu Leu Val Thr Lys Ala Val Lys Ala Gly Asp Ile Asp Ala 55 60 65 70 AAA ACC CGA CGT AGG ATC TTT GAT GCC GTT GGG TTC ACC TTT CCC AAC 652 Lys Thr Arg Arg Arg Ile Phe Asp Ala Val Gly Phe Thr Phe Pro Asn 75 80 85 CGA CTC CTG ACC ACT AAG GAG GCA CCT GAT GGC TGC CCT GAC CAC GTT 700 Arg Leu Leu Thr Thr Lys Glu Ala Pro Asp Gly Cys Pro Asp His Val 90 95 100 CTC CGG GCC CTG GGC GTG GCC CTG CTG GCC TGT TTC TGC AGC GAC CCT 748 Leu Arg Ala Leu Gly Val Ala Leu Leu Ala Cys Phe Cys Ser Asp Pro 105 110 115 GAA CTA GCC AGC CAT CCC CAG GTC CTG AAC AAG ATC CCC ATC CTT AGC 796 Glu Leu Ala Ser His Pro Gln Val Leu Asn Lys Ile Pro Ile Leu Ser 120 125 130 ACA TTC CTT ACA GCC CGG GGG GAC CCT GAT GAT GCT GCC CGC CGC TCC 844 Thr Phe Leu Thr Ala Arg Gly Asp Pro Asp Asp Ala Ala Arg A rg Ser 135 140 145 150 ATG ATC GAT GAC ACC TAC CAG TGC CTG ACA GCT GTT GCG GGC ACA CCC 892 Met Ile Asp Asp Thr Tyr Gln Cys Leu Thr Ala Val Ala Gly Thr Pro 155 160 165 CGA GGG CCC CGG CAC CTC ATT GCT GGT GGC ACC GTG TCT GCC CTG TGC 940 Arg Gly Pro Arg His Leu Ile Ala Gly Gly Thr Val Ser Ala Leu Cys 170 175 180 CAG GCG TAC CTG GGG CAT GGC TAC GGC TTT GAC CAG GCT CTG GCA CTC 988 Gln Ala Tyr Leu Gly His Gly Tyr Gly Phe Asp Gln Ala Leu Ala Leu 185 190 195 TTG GTG GGG CTG CTG GCT GCT GCA GAG ACA CAG TGC TGG AAG GAG GCG 1036 Leu Val Gly Leu Leu Ala Ala Ala Glu Thr Gln Cys Trp Lys Glu Ala 200 205 210 GAG CCC GAC CTG CTG GCT GTG TTG CGA GGC CTC AGT GAG GAT TTC CAA 1084 Glu Pro Asp Leu Leu Ala Val Leu Arg Gly Leu Ser Glu Asp Phe Gln 215 220 225 230 AGA GCC GAA GAT GCC AGC AAG TTT GAG CTC TGC CAG CTG CTG CCC CTT 1132 Arg Ala Glu Asp Ala Ser Lys Phe Glu Leu Cys Gln Leu Leu Pro Leu 235 240 245 TTT CTG CCC CCA ACA ACT GTG CCC CCT GAA TGC CAC CGA GAT CTG CAG 1180 Phe Leu Pro Pro Thr Thr Val Pro Pro G lu Cys His Arg Asp Leu Gln 250 255 260 GCT GGG CTG GCA CGA ATC CTA GGA AGC AAG TTG AGC TCC TGG CAG CGC 1228 Ala Gly Leu Ala Arg Ile Leu Gly Ser Lys Leu Ser Ser Trp Gln Arg 265 270 275 275 AAC CCT GCA CTG AAG CTG GCA GCC CGC CTG GCT CAT GCC TGC GGC TCC 1276 Asn Pro Ala Leu Lys Leu Ala Ala Arg Leu Ala His Ala Cys Gly Ser 280 285 290 GAC TGG ATC CCA GTG GGC AGC TCT GGG AGC AAG TTT CTG GCC CTG CTC 1324 Asp Trp Ile Pro Val Gly Ser Ser Gly Ser Lys Phe Leu Ala Leu Leu 295 300 305 310 GTG AAT CTA GCG TGC GTG GAG GTA CGG CTG GCT CTC GAG GAG ACA GGC 1372 Val Asn Leu Ala Cys Val Glu Val Arg Leu Ala Leu Glu Glu Thr Gly 315 320 325 ACA GAG GTG AAA GAA GAC GTG GTA ACA GCC TGC TAT GCC CTT ATG GAA 1420 Thr Glu Val Lys Glu Asp Val Val Thr Ala Cys Tyr Ala Leu Met Glu 330 335 340 TTG GGG ATC CAG GAG TGC ACC CGC TGT GAG CAG TCC CTG CTT AAG GAG 1468 Leu Gly Ile Gln Glu Cys Thr Arg Cys Glu Gln Ser Leu Leu Lys Glu 345 350 355 CCA CAG AAG GTT CAG CTC GTG AGC ATT ATG AAA GAG GCC ATT GGC GCT 1516 Pro Gln Lys Val Gl n Leu Val Ser Ile Met Lys Glu Ala Ile Gly Ala 360 365 370 GTC ATC CAC TAC CTG CTG CAG GTG GGG CCA GAG AAG CAG AAA GAG CCC 1564 Val Ile His Tyr Leu Leu Gln Val Gly Pro Glu Lys Gln Lys Glu Pro 375 380 385 390 TTT GTG TTT GCC TCG GTG CGG ATC CTG GGT GCC TGG CTG GCG GAG GAG 1612 Phe Val Phe Ala Ser Val Arg Ile Leu Gly Ala Trp Leu Ala Glu Glu 395 400 405 ACC ACC TCA TCC CTG CGT AAG GAG GTG TGC CAA CTG CTG CCC TTC CTT GTC 1660 Thr Ser Ser Leu Arg Lys Glu Val Cys Gln Leu Leu Pro Phe Leu Val 410 415 420 CGA TAT GCC AAG ACA CTC TAT GAG GAG GCT GAG GAG GCC AGT GAC ATT 1708 Arg Tyr Ala Lys Thr Leu Tyr Glu Glu Ala Glu Glu Ala Ser Asp Ile 425 430 435 TCG CAG CAG GTG GCC AAC TTG GCC ATC TCT CCT ACT ACA CCA GGG CCT 1756 Ser Gln Gln Val Ala Asn Leu Ala Ile Ser Pro Thr Thr Pro Gly Pro 440 445 450 TCA TGG CCA GGG GAT GCT CTC CGG CTC CTC CTT CCC GGC TGG TGT CAC 1804 Ser Trp Pro Gly Asp Ala Leu Arg Leu Leu Leu Pro Gly Trp Cys His 455 460 465 470 CTG ACT GTT GAA GAT GGT CCC CGG GAG ATT CTG ATC AAG GAA GGA GCC 1852 Leu Thr Val Glu Asp Gly Pro Arg Glu Ile Leu Ile Lys Glu Gly Ala 475 480 485 CCC TCA CTT CTG TGC AAG TAC TTC CTG CAG CAG TGG GAA CTC ACA TCC 1900 Pro Ser Leu Leu Cys Lys Tyr Phe Leu Gln Gln Trp Glu Leu Thr Ser 490 495 500 CCG GGC CAT GAT ACC TCG GTG CTG CCA GAC AGC GTG GAG ATC GGC CTT 1948 Pro Gly His Asp Thr Ser Val Leu Pro Asp Ser Val Glu Ile Gly Leu 505 510 515 CAG ACC TGT TGC CAC ATC TTC CTC AAC CTG GTG GTC ACC GCT CCA GGG 1996 Gln Thr Cys Cys His Ile Phe Leu Asn Leu Val Val Thr Ala Pro Gly 520 525 530 CTG ATC AAA CGC GAT GCC TGC TTC ACA TCT CTT ATG AAC ACC CTG ATG 2044 Leu Ile Lys Arg Asp Ala Cys Phe Thr Ser Leu Met Asn Thr Leu Met 535 540 545 550 ACG TCA CTG CCC TCA CTA GTG CAG CAA CAA GGG AGA CTG CTT CTA GCT 2092 Thr Ser Leu Pro Ser Leu Val Gln Gln Gln Gly Arg Leu Leu Leu Ala 555 560 565 GCC AAC GTG GCC ACT TTG GGG CTC CTA ATG GCC CGG CTC CTT AGC ACC 2140 Ala Asn Val Ala Thr Leu Gly Leu Leu Met Ala Arg Leu Leu Ser Thr 570 575 575 580 TCT CCA GCT CTC CAA GGA ACC CCA GCC TCC CGA GGT TTC TTC GCA GCT 2188 Ser Pro Ala Leu Gln Gly Thr Pro Ala Ser Arg Gly Phe Phe Ala Ala 585 590 595 GCC ATC CTC TTT CTG TCA CAG TCC CAT GTG GCA CGA GCC ACC CCT GGC 2236 Ala Ile Leu Phe Leu Ser Gln Ser His Val Ala Arg Ala Thr Pro Gly 600 605 610 TCT GAC CAG GCA GTG TTG GCC CTG TCC CCT GAC TAT GAA GGC ATC TGG 2284 Ser Asp Gln Ala Val Leu Ala Leu Ser Pro Asp Tyr Glu Gly Ile Trp 615 620 625 630 630 GCT GAC TTG CAA GAG CTC TGG TTC CTG GGC ATG CAG GCC TTC ACG GGT 2332 Ala Asp Leu Gln Glu Leu Trp Phe Leu Gly Met Gln Ala Phe Thr Gly 635 640 645 TGT GTG CCG CTG CTG CCC TGG CTG GCC CCT GCC GCC CTG CGC TCC 2380 Cys Val Pro Leu Leu Pro Trp Leu Ala Pro Ala Ala Leu Arg Ser Arg 650 655 660 TGG CCA CAG GAG CTG CTA CAA CTG CTA GGT AGT GTA AGC CCC AAC TCC 2428 Trp Pro Gln Glu Leu Leu Gln Leu Leu Gly Ser Val Ser Pro Asn Ser 665 670 675 GTC AAG CCT GAG ATG GTG GCT GCC TAC CAG GGC GTG CTG GTG GAA TTG 2476 Val Lys Pro Glu Met Val Ala Ala Tyr Gln Gly Val Leu Val Glu Leu 680 685 690 GCA CGG GCT AAC CGG CTA T GC CGG GAG GCC ATG AGG CTG CAG GCG GGT 2524 Ala Arg Ala Asn Arg Leu Cys Arg Glu Ala Met Arg Leu Gln Ala Gly 695 700 705 710 GAA GAA ACG GCC AGC CAT TAC CGA ATG GCT GCT TTG GAG CAG TGC CTG 2572 Glu Glu Thr Ala Ser His Tyr Arg Met Ala Ala Leu Glu Gln Cys Leu 715 720 725 TCA GAG CCC TGAGGGGCAT CCAGTGGGTA CAGACCCAGG CGGGCAGCGA 2621 Ser Glu Pro GGGAAGGAGG GAGGAGGCAT CTTCCCTGAA GCCCCCAAAC TGGACCCCTT CTTCAGACCC 2681 CCACAAACAC CCCAGCTTTC TGGCTTTTCT GAGGGCTAGG GCATGGTGCC CACCTCTCAA 2741 GTATAAGAAA CTGCATCCTG CCTCCAGCCC CCTTGGGGCA GGGATTGGCT TGGAACAGAG 2801 GTTGGCCCCG CCAGGCCGGG GAAGGTTGGA GAAGCCCCCA GGAGGAGGGC AACTAAGTGT 2861 CATTATACCC AGTGTCTGGC TCCCTGATAG GAGGGAGGTC CCAGGGTAGG AGCGGGCTGG 2921 CAGGCGCTGA CTGCCTCAGC CCATGTGCCC TGCCGGCCAG GGCGTGGCCT CCCCAAGGCT 2981 GTGGTGCCCC TTCTGGCTCC CCTAGGTCAG GTCCGCGCCC TTTAAATTGG CCGCTTGGCT 3041 TTTGCCTTTG GTCCTCTTGG ACAGAGAGCA GGCTCAGGCC ATTGACATCA CAGTTCTTCC 3101 TTTTAACTCT AGTGACCCGG GGTCCGAGTT GCCCCTATGC TTCCAGGGCA ATTTGGAGCA 3161 GACAGACCAG TGGGGGGT GG GGAACCTCCT TCCACCTGCG CTTCCTTGAG GGGACCAGAG 3221 AGCCCTTGGT CCCAGGTCTC TTGAGCTTTT GTGTCATGTT GCAGCAGAGT GAAGATGGGG 3281 GGTTGGGGGT TATTTATTTT GCTTGTCCTT ATCCCTGCTT GGACACCTGA GCATCAGATC 3341 CCTGTGCCCC TGGTGCCATC TGGCCTGCTG GAGCCAGGAA CAAGAGGTCA CCCCACCCTA 3401 GAATCCGCAT GGTTTCCCTG TGATTGCACT CCACTGCCAC CGTGGTGCCT GGCTTCAGCT 3461 CCCCTCCCCC AATCCCTGCT AAGCCTCTAC TCTGCAGGGA GACGCGACTG GCGGCTCCAG 3521 CAGGAACTAC CTTTCTGAAC CCGCGGAGAC CCGCATAAGC CTGACCCCTT GCTTCCTCCC 3581 CGCCCCCCAG TGCGTTCTGT GATCGCCAAG TTCAAAGCTG TGCACATGTG GACACTCAAT 3641 AAATGTTTAT TGGTGAAAAA AAAAAAAAAA AAA 3674

SEQ ID NO: 2 Sequence length: 729 Sequence type: amino acid Topology: Linear Sequence type: Peptide sequence Met Ser Cys Cys Asp Leu Ala Ala Ala Gly Gln Leu Gly Lys Ala Gly 5 10 15 Ile Met Ala Ser Asp Cys Glu Pro Ala Leu Asn Gln Ala Glu Ser Arg 20 25 30 Asn Pro Thr Leu Glu Arg Tyr Leu Gly Ala Leu Arg Glu Ala Lys Asn 35 40 45 Asp Ser Glu Gln Phe Ala Ala Leu Leu Leu Val Thr Lys Ala Val Lys 50 55 60 Ala Gly Asp Ile Asp Ala Lys Thr Arg Arg Arg Ile Phe Asp Ala Val 65 70 75 80 Gly Phe Thr Phe Pro Asn Arg Leu Leu Thr Thr Lys Glu Ala Pro Asp 85 90 95 Gly Cys Pro Asp His Val Leu Arg Ala Leu Gly Val Ala Leu Leu Ala 100 105 110 Cys Phe Cys Ser Asp Pro Glu Leu Ala Ser His Pro Gln Val Leu Asn 115 120 125 Lys Ile Pro Ile Leu Ser Thr Phe Leu Thr Ala Arg Gly Asp Pro Asp 130 135 140 Asp Ala Ala Arg Arg Ser Met Ile Asp Asp Thr Tyr Gln Cys Leu Thr 145 150 155 160 Ala Val Ala Gly Thr Pro Arg Gly Pro Arg His Leu Ile Ala Gly Gly 165 170 175 Thr Val Ser Ala Leu Cys Gln Ala Tyr Leu Gly His Gly Tyr Gly Phe 180 185 190 Asp Gln Ala Leu Ala Leu Leu Val Gly Leu Leu Ala Ala Ala Glu Thr 195 200 205 Gln Cys Trp Lys Glu Ala Glu Pro Asp Leu Leu Ala Val Leu Arg Gly 210 215 220 Leu Ser Glu Asp Phe Gln Arg Ala Glu Asp Ala Ser Lys Phe Glu Leu 225 230 235 240 Cys Gln Leu Leu Pro Leu Phe Leu Pro Pro Thr Thr Val Pro Pro Glu 245 250 255 Cys His Arg Asp Leu Gln Ala Gly Leu Ala Arg Ile Leu Gly Ser Lys 260 265 270 270 Leu Ser Ser Trp Gln Arg Asn Pro Ala Leu Lys Leu Ala Ala Arg Leu 275 280 285 Ala His Ala Cys Gly Ser Asp Trp Ile Pro Val Gly Ser Ser Gly Ser 290 295 300 Lys Phe Leu Ala Leu Leu Val Asn Leu Ala Cys Val Glu Val Arg Leu 305 310 315 320 Ala Leu Glu Glu Thr Gly Thr Glu Val Lys Glu Asp Val Val Thr Ala 325 330 335 Cys Tyr Ala Leu Met Glu Leu Gly Ile Gln Glu Cys Thr Arg Cys Glu 340 345 350 Gln Ser Leu Leu Lys Glu Pro Gln Lys Val Gln Leu Val Ser Ile Met 355 360 365 Lys Glu Ala Ile Gly Ala Val Ile His Tyr Leu Leu Gln Val Gly Pro 370 375 380 380 Glu Lys Gln Lys Glu Pro Phe Val Phe Ala Ser Val Arg Ile Leu Gly 385 390 395 400 400 Ala Trp Leu Ala Glu Glu Thr Ser Ser Leu Arg Lys Glu Val Cys Gln 405 410 415 Leu Leu Pro Phe Leu Val Arg Tyr Ala Lys Thr Leu Tyr Glu Glu Ala 420 425 430 Glu Glu Ala Ser Asp Ile Ser Gln Gln Val Ala Asn Leu Ala Ile Ser 435 440 445 Pro Thr Thr Pro Gly Pro Ser Trp Pro Gly Asp Ala Leu Arg Leu Leu 450 455 460 Leu Pro Gly Trp Cys His Leu Thr Val Glu Asp Gly Pro Arg Glu Ile 465 470 475 480 480 Leu Ile Lys Glu Gly Ala Pro Ser Leu Leu Cys Lys Tyr Phe Leu Gln 485 490 495 Gln Trp Glu Leu Thr Ser Pro Gly His Asp Thr Ser Val Leu Pro Asp 500 505 510 Ser Val Glu Ile Gly Leu Gln Thr Cys Cys His Ile Phe Leu Asn Leu 515 520 525 Val Val Thr Ala Pro Gly Leu Ile Lys Arg Asp Ala Cys Phe Thr Ser 530 535 540 Leu Met Asn Thr Leu Met Thr Ser Leu Pro Ser Leu Val Gln Gln Gln 545 550 555 560 Gly Arg Leu Leu Leu Ala Ala Asn Val Ala Thr Leu Gly Leu Leu Met 565 570 575 Ala Arg Leu Leu Ser Thr Ser Pro Ala Leu Gln Gly Thr Pro Ala Ser 580 585 590 Arg Gly Phe Phe Ala Ala Ala Ile Leu Phe Leu Ser Gln Ser His Val 595 600 605 Ala Arg Ala Thr Pro Gly Ser Asp Gln Ala Val Leu Ala Leu Ser Pro 610 615 620 620 Asp Tyr Glu Gly Ile Trp Ala Asp Leu Gln Glu Leu Trp Phe Leu Gly 625 630 635 640 Met Gln Ala Phe Thr Gly Cys Val Pro Leu Leu Pro Trp Leu Ala Pro 645 650 655 Ala Ala Leu Arg Ser Arg Trp Pro Gln Glu Leu Leu Gln Leu Leu Gly 660 665 670 Ser Val Ser Pro Asn Ser Val Lys Pro Glu Met Val Ala Ala Tyr Gln 675 680 685 Gly Val Leu Val Glu Leu Ala Arg Ala Asn Arg Leu Cys Arg Glu Ala 690 695 700 Met Arg Leu Gln Ala Gly Glu Glu Thr Ala Ser His Tyr Arg Met Ala 705 710 715 715 720 Ala Leu Glu Gln Cys Leu Ser Glu Pro 725

SEQ ID NO: 3 Sequence length: 3337 Sequence type: nucleic acid Number of strands: double-stranded Topology: linear Sequence type: cDNA Sequence characteristics Characteristic determination method: E sequence CGCGGCGTGA GCAGCAGCCC CAGGCTCCCGCGAGCATCGCG CCCGGAGAAA GACTTCGCCG 60 CTCGGGGCCG CAGCCTGGTG AGCTCAGCCG TTGGGCAAGG CGGGCATC ATG GCC TCG 117 Met Ala Ser GAT TGT GAG CCA GCT CTG AAC CAG GCA GAG AGC CGA AAC CCC ACC CTG 165 Asp Cys Glu Pro Ala Leu Asu Gln 15 GAG CGC TAC CTG GGA GCC CTC CGT GAG GCC AAG AAT GAC AGC GAG CAG 213 Glu Arg Tyr Leu Gly Ala Leu Arg Glu Ala Lys Asn Asp Ser Glu Gln 20 25 30 35 TTT GCA GCC CTG CTG CTA GTA ACC AAG GCA GTC AAA GCA GGT GAC ATT 261 Phe Ala Ala Leu Leu Leu Val Thr Lys Ala Val Lys Ala Gly Asp Ile 40 45 50 GAC GCC AAA ACC CGA CGT AGG ATC TTT GAT GCC GTT GGG TTC ACC TTT 309 Asp Ala Lys Thr Arg Arg Arg Ile Phe Asp Ala Val Gly Phe Thr Phe 55 60 65 CCC AAC CGA CTC CTG ACC ACT AAG GAG GCA CCT GAT GGC TGC CCT GAC 357 Pro Asn Arg Leu Leu Thr Thr Lys Glu Ala Pro Asp Gly Cys Pro Asp 70 75 80 CAC GTT CTC CGG GCC CTG GGC GTG GCC CTG CTG GCC TGT TTC TGC AGC 405 His Val Leu Arg Ala Leu Gly Val Ala Leu Leu Ala Cys Phe Cys Ser 85 90 95 GAC CCT GAA CTA GCC AGC CAT CCC CAG GTC CTG AAC AAG ATC CCC ATC 453 Asp Pro Glu Leu Ala Ser His Pro Gln Val Leu Asn Lys Ile Pro Ile 100 105 110 115 CTT AGC ACA TTC CTT ACA GCC CGG GGG GAC CCT GAT GAT GCT GCC CGC 501 Leu Ser Thr Phe Leu Thr Ala Arg Gly Asp Pro Asp Asp Ala Ala Arg 120 125 130 CGC TCC ATG ATC GAT GAC ACC TAC CAG TGC CTG ACA GCT GTT GCG GGC 549 Arg Ser Met Ile Asp Asp Thr Tyr Gln Cys Leu Thr Ala Val Ala Gly 135 140 145 ACA CCC CGA GGG CCC CGG CAC CTC ATT GCT GGT GGC ACC GTG TCT GCC 597 Thr Pro Arg Gly Pro Arg His Leu Ile Ala Gly Gly Thr Val Ser Ala 150 155 160 CTG TGC CAG GCG TAC CTG GGG CAT GGC TAC GGC TTT GAC CAG GCT CTG 645 Leu Cys Gln Ala Tyr Leu Gly His Gly Tyr Gly Phe Asp Gln Ala Leu 165 170 175 GCA CTC TTG GTG GGG CTG CTG GCT GCT GCA GAG ACA CAG TGC TGG AAG 693 Ala Leu Leu Val Gly Leu Leu Ala Ala Ala Glu Thr Gln Cys Trp Lys 180 185 190 195 GAG GCG GAG CCC GAC CTG CTG GCT GTG TTG CGA GGC CTC AGT GAG GAT 741 Glu Ala Glu Pro Asp Leu Leu Ala Val Leu Arg Gly Leu Ser Glu Asp 200 205 210 TTC CAA AGA GCC GAA GAT GCC AGC AAG TTT GAG CTC TGC CAG CTG CTG 789 Phe Gln Arg Ala Glu Asp Ala Ser Lys Phe Glu Leu Cys Gln Leu Leu 215 220 225 CCC CTT TTT CTG CCC CCA ACA ACT GTG CCC CCT GAA TGC CAC CGA GAT 837 Pro Leu Phe Leu Pro Pro Thr Thr Val Pro Pro Glu Cys His Arg Asp 230 235 240 CTG CAG GCT GGG CTG GCA CGA ATC CTA GGA AGC AAG TTG AGC TCC TGG 885 Leu Gln Ala Gly Leu Ala Arg Ile Leu Gly Ser Lys Leu Ser Ser Trp 245 250 255 CAG CGC AAC CCT GCA CTG AAG CTG GCA GCC CGC CTG GCT CAT GCC TGC 933 Gln Arg Asn Pro Ala Leu Lys Leu Ala Ala Arg Leu Ala His Ala Cys 260 265 270 275 275 GGC TCC GAC TGG ATC CCA GTG GGC AGC TCT GGG AGC AAG TTT CTG GCC 981 Gly Ser Asp Trp Ile Pro Val Gly Ser Ser Gly Ser Lys Phe Leu Ala 280 285 285 290 CTG CTC GTG AAT CTA GCG TGC GTGGAG GTA CGG CTG GCT CTC GAG GAG 1029 Leu Leu Val Asn Leu Ala Cys Val Glu Val Arg Leu Ala Leu Glu Glu 295 300 305 ACA GGC ACA GAG GTG AAA GAA GAC GTG GTA ACA GCC TGC TAT GCC CTT 1077 Thr Gly Thr Glu Val Lys Glu Asp Val Val Thr Ala Cys Tyr Ala Leu 310 315 320 ATG GAA TTG GGG ATC CAG GAG TGC ACC CGC TGT GAG CAG TCC CTG CTT 1125 Met Glu Leu Gly Ile Gln Glu Cys Thr Arg Cys Glu Gln Ser Leu Leu 325 330 335 AAG GAG CCA CAG AAG GTT CAG CTC GTG AGC ATT ATG AAA GAG GCC ATT 1173 Lys Glu Pro Gln Lys Val Gln Leu Val Ser Ile Met Lys Glu Ala Ile 340 345 350 355 GGC GCT GTC ATC CAC TAC CTG CTG CAG GTG GGG CCA GAG AAG CAG AAA 1221 Gly Ala Val Ile His Tyr Leu Leu Gln Val Gly Pro Glu Lys Gln Lys 360 365 370 GAG CCC TTT GTG TTT GCC TCG GTG CGG ATC CTG GGT GCC TGG CTG GCG 1269 Glu Pro Phe Val Phe Ala Ser Val Arg Ile Leu Gly Ala Trp Leu Ala 375 380 385 GAG GAG ACC TCA TCC CTG CGT AAG GAG GTG TGC CAA CTG CTG CCC TTC 1317 Glu Glu Glu Thr Ser Ser Leu Arg Lys Glu Val Cys Gln Leu Leu Pro Phe 390 395 400 CTT GTC CGA TAT GCC AAG ACA CTC TAT GAG GAG GCT GAG GAG GCC AGT 1365 Leu Val Arg Tyr Ala Lys Thr Leu Tyr Glu Glu Ala Glu Glu Ala Ser 405 410 415 GAC ATT TCG CAG CAG GTG GCC AAC TTG GCC ATC TCT CCT ACT ACA CCA 1413 Asp Ile Ser Gln Gln Val Ala Asn Leu Ala Ile Ser Pro Thr Thr Pro 420 425 430 435 GGG CCT TCA TGG CCA GGG GAT GCT CTC CGG CTC CTC CTT CCC GGC TGG 1461 Gly Pro Ser Trp Pro Gly Asp Ala Leu Arg Leu Leu Leu Pro Gly Trp 440 445 450 TGT CAC CTG ACT GTT GAA GAT GGT CCC CGG GAG ATT CTG ATC AAG GAA 1509 Cys His Leu Thr Val Glu Asp Gly Pro Arg Glu Ile Leu Ile Lys Glu 455 460 465 465 GGA GCC CCC TCA CTT CTG TGC AAG TAC TTC CTG CAG CAG TGG GAA CTC 1557 Gly Ala Pro Ser Leu Leu Cys Lys Tyr Phe Leu Gln Gln Trp Glu Leu 470 475 480 ACA TCC CCG GGC CAT GAT ACC TCG GTG CTG CCA GAC AGC GTG GAG ATC 1605 Thr Ser Pro Gly His Asp Thr Ser Val Leu Pro Asp Ser Val Glu Ile 485 490 495 GGC CTT CAG ACC TGT TGC CAC ATC TTC CTC AAC CTG GTG GTC ACC GCT 1653 Gly Leu Gln Thr Cys Cys His Ile Phe Leu Asn Leu Val Val Thr Ala 500 5 05 510 515 CCA GGG CTG ATC AAA CGC GAT GCC TGC TTC ACA TCT CTT ATG AAC ACC 1701 Pro Gly Leu Ile Lys Arg Asp Ala Cys Phe Thr Ser Leu Met Asn Thr 520 525 530 CTG ATG ACG TCA CTG CCC TCA CTA GTG CAG CAA CAA GGG AGA CTG CTT 1749 Leu Met Thr Ser Leu Pro Ser Leu Val Gln Gln Gln Gly Arg Leu Leu 535 540 545 CTA GCT GCC AAC GTG GCC ACT TTG GGG CTC CTA ATG GCC CGG CTC CTT 1797 Leu Ala Ala Asn Val Ala Thr Leu Gly Leu Leu Met Ala Arg Leu Leu 550 555 560 AGC ACC TCT CCA GCT CTC CAA GGA ACC CCA GCC TCC CGA GGT TTC TTC 1845 Ser Thr Ser Pro Ala Leu Gln Gly Thr Pro Ala Ser Arg Gly Phe Phe 565 570 575 GCA GCT GCC ATC CTC TTT CTG TCA CAG TCC CAT GTG GCA CGA GCC ACC 1893 Ala Ala Ala Ile Leu Phe Leu Ser Gln Ser His Val Ala Arg Ala Thr 580 585 590 590 CCT GGC TCT GAC CAG GCA GTG TTG GCC CTG TCC CCT GAC TAT GAA GGC 1941 Pro Gly Ser Asp Gln Ala Val Leu Ala Leu Ser Pro Asp Tyr Glu Gly 600 605 610 ATC TGG GCT GAC TTG CAA GAG CTC TGG TTC CTG GGC ATG CAG GCC TTC 1989 Ile Trp Ala Asp Leu Gln Glu Leu Trp Phe Leu Gl y Met Gln Ala Phe 615 620 625 ACG GGT TGT GTG CCG CTG CTG CCC TGG CTG GCC CCT GCC GCC CTG CGC 2037 Thr Gly Cys Val Pro Leu Leu Pro Trp Leu Ala Pro Ala Ala Leu Arg 630 635 640 TCC CGC TGG CCA CAG GAG CTG CTA CAA CTG CTA GGT AGT GTA AGC CCC 2085 Ser Arg Trp Pro Gln Glu Leu Leu Gln Leu Leu Gly Ser Val Ser Pro 645 650 655 655 AAC TCC GTC AAG CCT GAG ATG GTG GCT GCC TAC CAG GGC GTG CTG GTG 2133 Asn Ser Val Lys Pro Glu Met Val Ala Ala Tyr Gln Gly Val Leu Val 660 665 670 675 GAA TTG GCA CGG GCT AAC CGG CTA TGC CGG GAG GCC ATG AGG CTG CAG 2181 Glu Leu Ala Arg Ala Asn Arg Leu Cys Arg Glu Ala Met Arg Leu Gln 680 685 690 GCG GGT GAA GAA ACG GCC AGC CAT TAC CGA ATG GCT GCT TTG GAG CAG 2229 Ala Gly Glu Glu Thr Ala Ser His Tyr Arg Met Ala Ala Leu Glu Gln 695 700 705 TGC CTG TCA GAG CCC TGAGGGGCAT CCGATGGACAGACCA Leu Ser Glu Pro 710 GGGAAGGAGG GAGGAGGCAT CTTCCCTGAA GCCCCCAAAC TGGACCCCTT CTTCAGACCC 2344 CCACAAACAC CCCAGCTTTC TGGCTTTTCT GAGGGCTAGG GCATGGTGCC CACCTCTCAA 2404 GTATAA GAAA CTGCATCCTG CCTCCAGCCC CCTTGGGGCA GGGATTGGCT TGGAACAGAG 2464 GTTGGCCCCG CCAGGCCGGG GAAGGTTGGA GAAGCCCCCA GGAGGAGGGC AACTAAGTGT 2524 CATTATACCC AGTGTCTGGC TCCCTGATAG GAGGGAGGTC CCAGGGTAGG AGCGGGCTGG 2584 CAGGCGCTGA CTGCCTCAGC CCATGTGCCC TGCCGGCCAG GGCGTGGCCT CCCCAAGGCT 2644 GTGGTGCCCC TTCTGGCTCC CCTAGGTCAG GTCCGCGCCC TTTAAATTGG CCGCTTGGCT 2704 TTTGCCTTTG GTCCTCTTGG ACAGAGAGCA GGCTCAGGCC ATTGACATCA CAGTTCTTCC 2764 TTTTAACTCT AGTGACCCGG GGTCCGAGTT GCCCCTATGC TTCCAGGGCA ATTTGGAGCA 2824 GACAGACCAG TGGGGGGTGG GGAACCTCCT TCCACCTGCG CTTCCTTGAG GGGACCAGAG 2884 AGCCCTTGGT CCCAGGTCTC TTGAGCTTTT GTGTCATGTT GCAGCAGAGT GAAGATGGGG 2944 GGTTGGGGGT TATTTATTTT GCTTGTCCTT ATCCCTGCTT GGACACCTGA GCATCAGATC 3004 CCTGTGCCCC TGGTGCCATC TGGCCTGCTG GAGCCAGGAA CAAGAGGTCA CCCCACCCTA 3064 GAATCCGCAT GGTTTCCCTG TGATTGCACT CCACTGCCAC CGTGGTGCCT GGCTTCAGCT 3124 CCCCTCCCCC AATCCCTGCT AAGCCTCTAC TCTGCAGGGA GACGCGACTG GCGGCTCCAG 3184 CAGGAACTAC CTTTCTGAAC CCGCGGAGAC CCGCATAAGC CTGACCCCTT GCTTCCTCCC 3244 CGCCCCCCAG T GCGTTCTGT GATCGCCAAG TTCAAAGCTG TGCACATGTG GACACTCAAT 3304 AAATGTTTAT TGGTGAAAAA AAAAAAAAAA AAA 3337

SEQ ID NO: 4 Sequence Length: 712 Sequence Type: Amino Acid Topology: Linear Sequence Type: Peptide Sequence Met Ala Ser Asp Cys Glu Pro Ala Leu Asn Gln Ala Glu Ser Arg Asn 5 10 15 Pro Thr Leu Glu Arg Tyr Leu Gly Ala Leu Arg Glu Ala Lys Asn Asp 20 25 30 Ser Glu Gln Phe Ala Ala Leu Leu Leu Val Thr Lys Ala Val Lys Ala 35 40 45 Gly Asp Ile Asp Ala Lys Thr Arg Arg Arg Ile Phe Asp Ala Val Gly 50 55 60 Phe Thr Phe Pro Asn Arg Leu Leu Thr Thr Lys Glu Ala Pro Asp Gly 65 70 75 80 Cys Pro Asp His Val Leu Arg Ala Leu Gly Val Ala Leu Leu Ala Cys 85 90 95 Phe Cys Ser Asp Pro Glu Leu Ala Ser His Pro Gln Val Leu Asn Lys 100 105 110 Ile Pro Ile Leu Ser Thr Phe Leu Thr Ala Arg Gly Asp Pro Asp Asp 115 120 125 Ala Ala Arg Arg Ser Met Ile Asp Asp Thr Tyr Gln Cys Leu Thr Ala 130 135 140 Val Ala Gly Thr Pro Arg Gly Pro Arg His Leu Ile Ala Gly Gly Thr 145 150 155 160 Val Ser Ala Leu Cys Gln Ala Tyr Leu Gly His Gly Tyr Gly Phe Asp 165 170 175 Gln Ala Leu Ala Leu Leu Val Gly Leu Leu Ala Ala Ala Glu Thr Gln 180 185 190 Cys Trp Lys Glu Ala Glu Pro Asp Leu Leu Ala Val Leu Arg Gly Leu 195 200 205 Ser Glu Asp Phe Gln Arg Ala Glu Asp Ala Ser Lys Phe Glu Leu Cys 210 215 220 Gln Leu Leu Pro Leu Phe Leu Pro Pro Thr Thr Val Pro Pro Glu Cys 225 230 235 240 His Arg Asp Leu Gln Ala Gly Leu Ala Arg Ile Leu Gly Ser Lys Leu 245 250 255 Ser Ser Trp Gln Arg Asn Pro Ala Leu Lys Leu Ala Ala Arg Leu Ala 260 265 270 270 His Ala Cys Gly Ser Asp Trp Ile Pro Val Gly Ser Ser Gly Ser Lys 275 280 285 Phe Leu Ala Leu Leu Val Asn Leu Ala Cys Val Glu Val Arg Leu Ala 290 295 300 Leu Glu Glu Thr Gly Thr Glu Val Lys Glu Asp Val Val Thr Ala Cys 305 310 315 320 Tyr Ala Leu Met Glu Leu Gly Ile Gln Glu Cys Thr Arg Cys Glu Gln 325 330 335 Ser Leu Leu Lys Glu Pro Gln Lys Val Gln Leu Val Ser Ile Met Lys 340 345 350 Glu Ala Ile Gly Ala Val Ile His Tyr Leu Leu Gln Val Gly Pro Glu 355 360 365 Lys Gln Lys Glu Pro Phe Val Phe Ala Ser Val Arg Ile Leu Gly Ala 370 375 380 380 Trp Leu Ala Glu Glu Thr Ser Ser Leu Arg Lys Glu Val Cys Gln Leu 385 390 395 400 Leu Pro Phe Leu Val Arg Tyr Ala Lys Thr Leu Tyr Glu Glu Ala Glu 405 410 415 Glu Ala Ser Asp Ile Ser Gln Gln Val Ala Asn Leu Ala Ile Ser Pro 420 425 430 Thr Thr Pro Gly Pro Ser Trp Pro Gly Asp Ala Leu Arg Leu Leu Leu 435 440 445 Pro Gly Trp Cys His Leu Thr Val Glu Asp Gly Pro Arg Glu Ile Leu 450 455 460 Ile Lys Glu Gly Ala Pro Ser Leu Leu Cys Lys Tyr Phe Leu Gln Gln 465 470 475 480 Trp Glu Leu Thr Ser Pro Gly His Asp Thr Ser Val Leu Pro Asp Ser 485 490 495 Val Glu Ile Gly Leu Gln Thr Cys Cys His Ile Phe Leu Asn Leu Val 500 505 510 Val Thr Ala Pro Gly Leu Ile Lys Arg Asp Ala Cys Phe Thr Ser Leu 515 520 525 Met Asn Thr Leu Met Thr Ser Leu Pro Ser Leu Val Gln Gln Gln Gly 530 535 535 540 Arg Leu Leu Leu Ala Ala Asn Val Ala Thr Leu Gly Leu Leu Met Ala 545 550 555 560 Arg Leu Leu Ser Thr Ser Pro Ala Leu Gln Gly Thr Pro Ala Ser Arg 565 570 575 Gly Phe Phe Ala Ala Ala Ile Leu Phe Leu Ser Gln Ser His Val Ala 580 585 590 Arg Ala Thr Pro Gl y Ser Asp Gln Ala Val Leu Ala Leu Ser Pro Asp 595 600 605 Tyr Glu Gly Ile Trp Ala Asp Leu Gln Glu Leu Trp Phe Leu Gly Met 610 615 620 Gln Ala Phe Thr Gly Cys Val Pro Leu Leu Pro Trp Leu Ala Pro Ala 625 630 635 640 Ala Leu Arg Ser Arg Trp Pro Gln Glu Leu Leu Gln Leu Leu Gly Ser 645 650 655 Val Ser Pro Asn Ser Val Lys Pro Glu Met Val Ala Ala Tyr Gln Gly 660 665 670 Val Leu Val Glu Leu Ala Arg Ala Asn Arg Leu Cys Arg Glu Ala Met 675 680 685 Arg Leu Gln Ala Gly Glu Glu Thr Ala Ser His Tyr Arg Met Ala Ala 690 695 700 Leu Glu Gln Cys Leu Ser Glu Pro 705 710

SEQ ID NO: 5 Sequence length: 2293 Sequence type: nucleic acid Number of strands: double-stranded Topology: linear Sequence type: cDNA Sequence characteristics Characteristic determination method: E sequence GGCGTGAGCA GCGGCCCGAG GCTCCCGGAG CATCGCGCTG GGAGAAGACT TCGCCGCTCG 60 GGGCCGCAGC CTGTGATCTC ATCGCCGCCC TGTCGTGACT TCATCA ATG TCG TGT 115 Met Ser Cys TGT GAC CTG GCT GCG GCG GGA CAG TTG GGC AAG GCG AGC ATC ATG GCC 163 Cys Asp Leu Ala Ala Gla Gly Aln 15 TCG GAT TGC GAG CCA GCT CTG AAC CAG GCA GAG GGC CGA AAC CCC ACC 211 Ser Asp Cys Glu Pro Ala Leu Asn Gln Ala Glu Gly Arg Asn Pro Thr 20 25 30 35 CTG GAG CGC TAC CTG GGA GCC CTC CGT GAG GCC AAG AAT GAC AGC GAG 259 Leu Glu Arg Tyr Leu Gly Ala Leu Arg Glu Ala Lys Asn Asp Ser Glu 40 45 50 CAG TTT GCA GCC CTG CTG CTA GTG ACC AAG GCA GTC AAA GCA GGT GAC 307 Gln Phe Ala Ala Lela Leu Leu Leu Val Thr Lys Ala Val Lys Ala Gly Asp 55 60 65 ATA GAT GCC AAA ACT CGG CGG CGG ATC TTC GAT GCT GTC GGC T TC ACC 355 Ile Asp Ala Lys Thr Arg Arg Arg Ile Phe Asp Ala Val Gly Phe Thr 70 75 80 TTC CCC AAT CGT CTC CTG ACC ACC AAG GAG GCG CCG GAT GGC TGC CCT 403 Phe Pro Asn Arg Leu Leu Thr Thr Lys Glu Ala Pro Asp Gly Cys Pro 85 90 95 GAC CAT GTT CTG CGG GCT TTG GGT GTG GCC CTG CTG GCC TGC TTC TGC 451 Asp His Val Leu Arg Ala Leu Gly Val Ala Leu Leu Ala Cys Phe Cys 100 105 110 115 AGT GAC CCT GAA CTG GCC GCC CAT CCC CAA GTC CTG AAC AAG ATT CCC 499 Ser Asp Pro Glu Leu Ala Ala His Pro Gln Val Leu Asn Lys Ile Pro 120 125 130 ATT CTT AGC ACC TTC CTC ACA GCC CGG GGG GAC CCG GAC GAT GCT GCC 547 Ile Leu Ser Thr Phe Leu Thr Ala Arg Gly Asp Pro Asp Asp Ala Ala 135 140 145 CGC CGC TCC ATG ATT GAT GAC ACC TAC CAG TGC CTG ACA GCT GTT GCG 595 Arg Arg Ser Met Ile Asp Asp Thr Tyr Gln Cys Leu Thr Ala Val Ala 150 155 160 GGC ACA CCC CGA GGG CCC CGG CAC CTC ATT GCT GGT GGC ACC GTG TCT 643 Gly Thr Pro Arg Gly Pro Arg His Leu Ile Ala Gly Gly Thr Val Ser 165 170 175 GCC CTG TGC CAG GCG TAC CTG GGG CAT GGC TAC GGC TT T GAC CAG GCT 691 Ala Leu Cys Gln Ala Tyr Leu Gly His Gly Tyr Gly Phe Asp Gln Ala 180 185 190 195 CTG GCA CTC TTG GTG GGG CTG CTG GCT GCT GCA GAG ACA CAG TGC TGG 739 Leu Ala Leu Leu Val Gly Leu Leu Ala Ala Ala Glu Thr Gln Cys Trp 200 205 210 AAG GAG GCG GAG CCC GAC CTG CTG GCT GTG TTG CGA GGC CTC AGT GAG 787 Lys Glu Ala Glu Pro Asp Leu Leu Ala Val Leu Arg Gly Leu Ser Glu 215 220 225 GAT TTC CAG AAA GCC GAG GAT GCC AGC AAG TTT GAG CTC TGC CAG CTG 835 Asp Phe Gln Lys Ala Glu Asp Ala Ser Lys Phe Glu Leu Cys Gln Leu 230 235 240 CTG CCC CTT TTT CTG CCC CCA ACA ACT GTG CCC CCT GAA TGC CAC CGA 883 Leu Pro Leu Phe Leu Pro Pro Thr Thr Val Pro Pro Glu Cys His Arg 245 250 255 GAT CTG CAG GCT GGG CTG GCA CGA ATC CTA GGA AGC AAG TTG AGC TCC 931 Asp Leu Gln Ala Gly Leu Ala Arg Ile Leu Gly Ser Lys Leu Ser Ser 260 265 270 275 TGG CAG CGC AAC CCT GCA CTG AAG CTG GCA GCC CGC CTG GCT CAT GCC 979 Trp Gln Arg Asn Pro Ala Leu Lys Leu Ala Ala Arg Leu Ala His Ala 280 285 290 290 TGC GGC TCC GAC TGG ATC CCA GTG GG C AGC TCT GGG AGC AAG TTT CTG 1027 Cys Gly Ser Asp Trp Ile Pro Val Gly Ser Ser Gly Ser Lys Phe Leu 295 300 305 GCC CTG CTC GTG AAT CTG GCG TGC GTG GAG GTA CGG CTG GCT CTC GAG 1075 Ala Leu Leu Val Asn Leu Ala Cys Val Glu Val Arg Leu Ala Leu Glu 310 315 320 GAG ACA AGC ACA GAG GTG AAA GAA GAC GTG GTA ACA GCC TGC TAT GCC 1123 Glu Thr Ser Thr Glu Val Lys Glu Asp Val Val Thr Ala Cys Tyr Ala 325 330 335 CTT ATG GAA TTG GGG ATC CAG GAG TGC ACC CGC TGT GAG CAG TCC CTG 1171 Leu Met Glu Leu Gly Ile Gln Glu Cys Thr Arg Cys Glu Gln Ser Leu 340 345 350 355 CTT AAG GAG CCA CAG AAG GTT CAG CTC GTG AGC ATT ATG AAA GAG GCC 1219 Leu Lys Glu Pro Gln Lys Val Gln Leu Val Ser Ile Met Lys Glu Ala 360 365 370 ATT GGC GCT GTC ATC CAC TAC CTG CTG CAG GTG GGG CCA GAG AAG CAG 1267 Ile Gly Ala Val Ile His Tyr Leu Leu Gln Val Gly Pro Glu Lys Gln 375 380 385 AAA GAG CCC TTT GTG TTT GCC TCG GTG CGG ATC CTG GGT GCC TGG CTG 1315 Lys Glu Pro Phe Val Phe Ala Ser Val Arg Ile Leu Gly Ala Trp Leu 390 395 400 GCG GAG GAG AC C TCA TCC CTG CGT AAG GAG GTG TGC CAA CTG CTG CCC 1363 Ala Glu Glu Thr Ser Ser Leu Arg Lys Glu Val Cys Gln Leu Leu Pro 405 410 415 TTC CTT GTC CGA TAT GCC AAG ACA CTC TAT GGG GAG GCT GAG GAG GCC 1411 Phe Leu Val Arg Tyr Ala Lys Thr Leu Tyr Gly Glu Ala Glu Glu Ala 420 425 430 435 AGT GAC ATT TCG CAG CAG GTG GCC AAC TTG GCC ATC TCT CCT ACT ACA 1459 Ser Asp Ile Ser Gln Gln Val Ala Asn Leu Ala Ile Ser Pro Thr Thr 440 445 450 CCA GGG CCT TCA TGG CCA GGG GAT GCT CTC CGG CTC CTC CTT CCC GGC 1507 Pro Gly Pro Ser Trp Pro Gly Asp Ala Leu Arg Leu Leu Leu Pro Gly 455 460 465 465 TGG TGT CAC CTG ACT GTT GAA GAT GGT CCC CGG GAG ATT CTG ATC AAG 1555 Trp Cys His Leu Thr Val Glu Asp Gly Pro Arg Glu Ile Leu Ile Lys 470 475 480 GAA GGA GCC CCC TCA CTT CTG TGC AAG TAC TTC CTG CAG CAG TGG GAA 1603 Glu Gly Ala Pro Ser Leu Leu Cys Lys Tyr Phe Leu Gln Gln Trp Glu 485 490 495 CTC ACA TCC CCG GGC CAT GAT ACC TCG GTG CTG CCA GAC AGC GTG GAG 1651 Leu Thr Ser Pro Gly His Asp Thr Ser Val Leu Pro Asp Ser Val Glu 500 505 510 515 ATC GGC CTT CAG ACC TGT TGC CAC ATC TTC CTC AAC CTG GTG GTC ACC 1699 Ile Gly Leu Gln Thr Cys Cys His Ile Phe Leu Asn Leu Val Val Thr 520 525 530 530 GCT CCA GGG CTG ATC AAA CGC GAT GCC TGC TTC ACA TCT CTT ATG AAC 1747 Ala Pro Gly Leu Ile Lys Arg Asp Ala Cys Phe Thr Ser Leu Met Asn 535 540 545 ACC CTG ATG ACG TCA CTG CCC TCA CTA GTG CAG CAA CAA GGG AGA CTG 1795 Thr Leu Met Thr Ser Leu Pro Ser Leu Val Gln Gln Gln Gly Arg Leu 550 555 560 CTT CTA GCT GCC AAC GTG GCC ACT TTG GGG CTC CTA ATG GCC CGG CTC 1843 Leu Leu Ala Ala Asn Val Ala Thr Leu Gly Leu Leu Met Ala Arg Leu 565 570 570 575 CTT AGC ACCCT CCA GCT CTC CAA GGA ACC CCA GCC TCC CGA GGT TTC 1891 Leu Ser Thr Ser Pro Ala Leu Gln Gly Thr Pro Ala Ser Arg Gly Phe 580 585 590 595 TTC GCA GCT GCC ATC CTC TTT CTG TCA CAG TCC CAT GTG GCA CGA GCC 1939 Phe Ala Ala Ala Ile Leu Phe Leu Ser Gln Ser His Val Ala Arg Ala 600 605 610 ACC CCT GGC TCT GAC CAG GCA GTG TTG GCC CTG TCC CCT GAC TAT GAA 1987 Thr Pro Gly Ser Asp Gln Ala Val Leu Ala Leu Ser Pro Asp Tyr Glu 615 620 625 GGC ATC TGG GCT GAC TTG CAA GAG CTC TGG TTC CTG GGC ATG CAG GCC 2035 Gly Ile Trp Ala Asp Leu Gln Glu Leu Trp Phe Leu Gly Met Gln Ala 630 635 635 640 TTC ACG GGT TGT GTG CC CTG CCC TGG CTG GCC CCT GCC GCC CTG 2083 Phe Thr Gly Cys Val Pro Leu Leu Pro Trp Leu Ala Pro Ala Ala Leu 645 650 655 655 CGC TCC CGC TGG CCA CAG GAG CTG CTA CAA CTG CTA GGT AGT GTA AGC 2131 Arg Ser Arg Trp Pro Gln Glu Leu Leu Gln Leu Leu Gly Ser Val Ser 660 665 670 675 CCC AAC TCC GTC AAG CCT GAG ATG GTG GCT GCC TAC CAG GGC GTG CTG 2179 Pro Asn Ser Val Lys Pro Glu Met Val Ala Ala Tyr Gln Gly Val Leu 680 685 690 GTG GAA TTG GCA CGG GCT AAC CGG CTA TGC CGG GAG GCC ATG AGG CTG 2227 Val Glu Leu Ala Arg Ala Asn Arg Leu Cys Arg Glu Ala Met Arg Leu 695 700 705 CAG GCG GGT GAA GAA ACG GCC AGC CAT TAC CGA AGC GCT GCT TTG GAG 2275 Gln Ala Gly Glu Glu Thr Ala Ser His Tyr Arg Met Ala Ala Leu Glu 710 715 720 CAG TGC CTG TCA GAG CCC 2293 Gln Cys Leu Ser Glu Pro 725

SEQ ID NO: 6 Sequence Length: 729 Sequence Type: Amino Acid Topology: Linear Sequence Type: Peptide Sequence Met Ser Cys Cys Asp Leu Ala Ala Ala Gly Gln Leu Gly Lys Ala Ser 5 10 15 Ile Met Ala Ser Asp Cys Glu Pro Ala Leu Asn Gln Ala Glu Gly Arg 20 25 30 Asn Pro Thr Leu Glu Arg Tyr Leu Gly Ala Leu Arg Glu Ala Lys Asn 35 40 45 Asp Ser Glu Gln Phe Ala Ala Leu Leu Leu Val Thr Lys Ala Val Lys 50 55 60 Ala Gly Asp Ile Asp Ala Lys Thr Arg Arg Arg Ile Phe Asp Ala Val 65 70 75 80 Gly Phe Thr Phe Pro Asn Arg Leu Leu Thr Thr Lys Glu Ala Pro Asp 85 90 95 Gly Cys Pro Asp His Val Leu Arg Ala Leu Gly Val Ala Leu Leu Ala 100 105 110 Cys Phe Cys Ser Asp Pro Glu Leu Ala Ala His Pro Gln Val Leu Asn 115 120 125 Lys Ile Pro Ile Leu Ser Thr Phe Leu Thr Ala Arg Gly Asp Pro Asp 130 135 140 Asp Ala Ala Arg Arg Ser Met Ile Asp Asp Thr Tyr Gln Cys Leu Thr 145 150 155 160 Ala Val Ala Gly Thr Pro Arg Gly Pro Arg His Leu Ile Ala Gly Gly 165 170 175 Thr Val Ser Ala Leu Cys Gln Ala Tyr Leu Gly His Gly Tyr Gly Phe 180 185 190 Asp Gln Ala Leu Ala Leu Leu Val Gly Leu Leu Ala Ala Ala Glu Thr 195 200 205 Gln Cys Trp Lys Glu Ala Glu Pro Asp Leu Leu Ala Val Leu Arg Gly 210 215 220 Leu Ser Glu Asp Phe Gln Lys Ala Glu Asp Ala Ser Lys Phe Glu Leu 225 230 235 240 Cys Gln Leu Leu Pro Leu Phe Leu Pro Pro Thr Thr Val Pro Pro Glu 245 250 255 Cys His Arg Asp Leu Gln Ala Gly Leu Ala Arg Ile Leu Gly Ser Lys 260 265 270 270 Leu Ser Ser Trp Gln Arg Asn Pro Ala Leu Lys Leu Ala Ala Arg Leu 275 280 285 Ala His Ala Cys Gly Ser Asp Trp Ile Pro Val Gly Ser Ser Gly Ser 290 295 300 Lys Phe Leu Ala Leu Leu Val Asn Leu Ala Cys Val Glu Val Arg Leu 305 310 315 320 Ala Leu Glu Glu Thr Ser Thr Glu Val Lys Glu Asp Val Val Thr Ala 325 330 335 Cys Tyr Ala Leu Met Glu Leu Gly Ile Gln Glu Cys Thr Arg Cys Glu 340 345 350 Gln Ser Leu Leu Lys Glu Pro Gln Lys Val Gln Leu Val Ser Ile Met 355 360 365 Lys Glu Ala Ile Gly Ala Val Ile His Tyr Leu Leu Gln Val Gly Pro 370 375 380 380 Glu Lys Gln Lys Glu Pro Phe Val Phe Ala Ser Val Arg Ile Leu Gly 385 390 395 400 400 Ala Trp Leu Ala Glu Glu Thr Ser Ser Leu Arg Lys Glu Val Cys Gln 405 410 415 Leu Leu Pro Phe Leu Val Arg Tyr Ala Lys Thr Leu Tyr Gly Glu Ala 420 425 430 Glu Glu Ala Ser Asp Ile Ser Gln Gln Val Ala Asn Leu Ala Ile Ser 435 440 445 Pro Thr Thr Pro Gly Pro Ser Trp Pro Gly Asp Ala Leu Arg Leu Leu 450 455 460 Leu Pro Gly Trp Cys His Leu Thr Val Glu Asp Gly Pro Arg Glu Ile 465 470 475 480 480 Leu Ile Lys Glu Gly Ala Pro Ser Leu Leu Cys Lys Tyr Phe Leu Gln 485 490 495 Gln Trp Glu Leu Thr Ser Pro Gly His Asp Thr Ser Val Leu Pro Asp 500 505 510 Ser Val Glu Ile Gly Leu Gln Thr Cys Cys His Ile Phe Leu Asn Leu 515 520 525 Val Val Thr Ala Pro Gly Leu Ile Lys Arg Asp Ala Cys Phe Thr Ser 530 535 540 Leu Met Asn Thr Leu Met Thr Ser Leu Pro Ser Leu Val Gln Gln Gln 545 550 555 560 Gly Arg Leu Leu Leu Ala Ala Asn Val Ala Thr Leu Gly Leu Leu Met 565 570 575 Ala Arg Leu Leu Ser Thr Ser Pro Ala Leu Gln Gly Thr Pro Ala Ser 580 585 590 Arg Gly Phe Phe Al a Ala Ala Ile Leu Phe Leu Ser Gln Ser His Val 595 600 605 Ala Arg Ala Thr Pro Gly Ser Asp Gln Ala Val Leu Ala Leu Ser Pro 610 615 620 Asp Tyr Glu Gly Ile Trp Ala Asp Leu Gln Glu Leu Trp Phe Leu Gly 625 630 635 640 Met Gln Ala Phe Thr Gly Cys Val Pro Leu Leu Pro Trp Leu Ala Pro 645 650 655 Ala Ala Leu Arg Ser Arg Trp Pro Gln Glu Leu Leu Gln Leu Leu Gly 660 665 670 670 Ser Val Ser Pro Asn Ser Val Lys Pro Glu Met Val Ala Ala Tyr Gln 675 680 685 Gly Val Leu Val Glu Leu Ala Arg Ala Asn Arg Leu Cys Arg Glu Ala 690 695 700 Met Arg Leu Gln Ala Gly Glu Glu Thr Ala Ser His Tyr Arg Met Ala 705 710 710 715 715 720 Ala Leu Glu Gln Cys Leu Ser Glu Pro 725

SEQ ID NO: 7 Sequence length: 2301 Sequence type: nucleic acid Number of strands: double-stranded Topology: linear Sequence type: cDNA Sequence characteristics Characteristic determination method: E sequence CGCGGCGTGA GCAGCGGCCC GAGGCTCCCGCGAGCATCGCG CTGGGAGAAG ACTTCGCCGC 60 TCGGGGCCGC AGCCTGTTGG GCAAGGCGAG CATC ATG GCC TCG GAT TGC GAG 112 Met Ala Ser Asp Cys Glu 5 CCA GCT CTG AAC CAG GCA GAG GGC CGA AAC CCC ACC CTG GAG CGC TAC 160 Pro Ala Leu Asn Gln Agn Gln As Leu Glu Arg Tyr 10 15 20 CTG GGA GCC CTC CGT GAG GCC AAG AAT GAC AGC GAG CAG TTT GCA GCC 208 Leu Gly Ala Leu Arg Glu Ala Lys Asn Asp Ser Glu Gln Phe Ala Ala 25 30 35 CTG CTG CTA GTG ACC AAG GCA GTC AAA GCA GGT GAC ATA GAT GCC AAA 256 Leu Leu Leu Val Thr Lys Ala Val Lys Ala Gly Asp Ile Asp Ala Lys 40 45 50 ACT CGG CGG CGG ATC TTC GAT GCT GTC GGC TTC ACC TTC CCC AAT CGT 304 Thr Arg Arg Arg Ile Phe Asp Ala Val Gly Phe Thr Phe Pro Asn Arg 55 60 65 70 CTC CTG ACC ACC AAG GAG GCG CCG GAT GGC TG C CCT GAC CAT GTT CTG 352 Leu Leu Thr Thr Lys Glu Ala Pro Asp Gly Cys Pro Asp His Val Leu 75 80 85 CGG GCT TTG GGT GTG GCC CTG CTG GCC TGC TTC TGC AGT GAC CCT GAA 400 Arg Ala Leu Gly Val Ala Leu Leu Ala Cys Phe Cys Ser Asp Pro Glu 90 95 100 CTG GCC GCC CAT CCC CAA GTC CTG AAC AAG ATT CCC ATT CTT AGC ACC 448 Leu Ala Ala His Pro Gln Val Leu Asn Lys Ile Pro Ile Leu Ser Thr 105 110 115 TTC CTC ACA GCC CGG GGG GAC CCG GAC GAT GCT GCC CGC CGC TCC ATG 496 Phe Leu Thr Ala Arg Gly Asp Pro Asp Asp Ala Ala Arg Arg Ser Met 120 125 130 ATT GAT GAC ACC TAC CAG TGC CTG ACA GCT GTT GCG GGC ACA CCC CGA 544 Ile Asp Asp Thr Tyr Gln Cys Leu Thr Ala Val Ala Gly Thr Pro Arg 135 140 145 150 GGG CCC CGG CAC CTC ATT GCT GGT GGC ACC GTG TCT GCC CTG TGC CAG 592 Gly Pro Arg His Leu Ile Ala Gly Gly Thr Val Ser Ala Leu Cys Gln 155 160 165 GCG TAC CTG GGG CAT GGC TAC GGC TTT GAC CAG GCT CTG GCA CTC TTG 640 Ala Tyr Leu Gly His Gly Tyr Gly Phe Asp Gln Ala Leu Ala Leu Leu 170 175 180 GTG GGG CTG CTG GCT GCT GCA GAG ACA CAG TGC TGG AAG GAG GCG GAG 688 Val Gly Leu Leu Ala Ala Ala Glu Thr Gln Cys Trp Lys Glu Ala Glu 185 190 195 CCC GAC CTG CTG GCT GTG TTG CGA GGC CTC AGT GAG GAT TTC CAG AAA 736 Pro Asp Leu Leu Ala Val Leu Arg Gly Leu Ser Glu Asp Phe Gln Lys 200 205 210 GCC GAG GAT GCC AGC AAG TTT GAG CTC TGC CAG CTG CTG CCC CTT TTT 784 Ala Glu Asp Ala Ser Lys Phe Glu Leu Cys Gln Leu Leu Pro Leu Phe 215 220 225 230 CTG CCC CCA ACA ACT GTG CCC CCT GAA TGC CAC CGA GAT CTG CAG GCT 832 Leu Pro Pro Thr Thr Val Val Pro Glu Cys His Arg Asp Leu Gln Ala 235 240 245 GGG CTG GCA CGA ATC CTA GGA AGC AAG TTG AGC TCC TGG CAG CGC AAC 880 Gly Leu Ala Arg Ile Leu Gly Ser Lys Leu Ser Ser Trp Gln Arg Asn 250 255 260 CCT GCA CTG AAG CTG GCA GCC CGC CTG GCT CAT GCC TGC GGC TCC GAC 928 Pro Ala Leu Lys Leu Ala Ala Arg Leu Ala His Ala Cys Gly Ser Asp 265 270 275 TGG ATC CCA GTG GGC AGC TCT GGG AGC AAG TTT CTG GCC CTG CTC GTG 976 Trp Ile Pro Val Gly Ser Ser Gly Ser Lys Phe Leu Ala Leu Leu Val 280 285 290 AAT CTG GCG TGC GTG GAG GTA CGG CTG GCT CTC GAG GAG ACA AGC ACA 1024 Asn Leu Ala Cys Val Glu Val Arg Leu Ala Leu Glu Glu Thr Ser Thr 295 300 305 310 GAG GTG AAA GAA GAC GTG GTA ACA GCC TGC TAT GCC CTT ATG GAA TTG 1072 Glu Val Lys Glu Asp Val Val Thr Ala Cys Tyr Ala Leu Met Glu Leu 315 320 325 GGG ATC CAG GAG TGC ACC CGC TGT GAG CAG TCC CTG CTT AAG GAG CCA 1120 Gly Ile Gln Glu Cys Thr Arg Cys Glu Gln Ser Leu Leu Lys Glu Pro 330 335 340 CAG AAG GTT CAG CTC GTG AGC ATT ATG AAA GAG GCC ATT GGC GCT GTC 1168 Gln Lys Val Gln Leu Val Ser Ile Met Lys Glu Ala Ile Gly Ala Val 345 350 355 ATC CAC TAC CTG CTG CAG GTG GGG CCA GAG AAG CAG AAA GAG CCC TTT 1216 Ile His Tyr Leu Leu Gln Val Gly Pro Glu Lys Gln Lys Glu Pro Phe 360 365 370 GTG TTT GCC TCG GTG CGG ATC CTG GGT GCC TGG CTG GCG GAG GAG ACC 1264 Val Phe Ala Ser Val Arg Ile Leu Gly Ala Trp Leu Ala Glu Glu Thr 375 380 385 390 TCA TCC CTG CGT AAG GAG GTG TGC CAA CTG CTG CCC TTC CTT GTC CGA 1312 Ser Ser Leu Arg Lys Glu Val Cys Gln Leu Leu Pro Phe Leu Val Arg 395 400 405 TAT GCC AAG ACA CTC TAT GGG GAG GCT GAG GAG GCC AGT GAC ATT TCG 1360 Tyr Ala Lys Thr Leu Tyr Gly Glu Ala Glu Glu Ala Ser Asp Ile Ser 410 415 420 CAG CAG GTG GCC AAC TTG GCC ATC TCT CCT ACT ACA CCA GGG CCT TCA 1408 Gln Gln Val Ala Asn Leu Ala Ile Ser Pro Thr Thr Pro Gly Pro Ser 425 430 435 TGG CCA GGG GAT GCT CTC CGG CTC CTC CTT CCC GGC TGG TGT CAC CTG 1456 Trp Pro Gly Asp Ala Leu Arg Leu Leu Leu Pro Gly Trp Cys His Leu 440 445 450 ACT GTT GAA GAT GGT CCC CGG GAG ATT CTG ATC AAG GAA GGA GCC CCC 1504 Thr Val Glu Asp Gly Pro Arg Glu Ile Leu Ile Lys Glu Gly Ala Pro 455 460 465 470 470 TCA CTT CTG TGC AAG TAC TTC CTG CAG CAG TGG GAA CTC ACA TCC CCG 1552 Ser Leu Leu Cys Lys Tyr Phe Leu Gln Gln Trp Glu Leu Thr Ser Pro 475 480 485 GGC CAT GAT ACC TCG GTG CTG CCA GAC AGC GTG GAG ATC GGC CTT CAG 1600 Gly His Asp Thr Ser Val Leu Pro Asp Ser Val Glu Ile Gly Leu Gln 490 495 500 ACC TGT TGC CAC ATC TTC CTC AAC CTG GTG GTC ACC GCT CCA GGG CTG 1648 Thr Cys Cys His Ile Phe Leu Asn Leu Val Val Thr Ala Pro G ly Leu 505 510 515 ATC AAA CGC GAT GCC TGC TTC ACA TCT CTT ATG AAC ACC CTG ATG ACG 1696 Ile Lys Arg Asp Ala Cys Phe Thr Ser Leu Met Asn Thr Leu Met Thr 520 525 530 530 TCA CTG CCC TCA CTA GTG CAG CAA CAA GGG AGA CTG CTT CTA GCT GCC 1744 Ser Leu Pro Ser Leu Val Gln Gln Gln Gly Arg Leu Leu Leu Ala Ala 535 540 545 550 550 AAC GTG GCC ACT TTG GGG CTC CTA ATG GCC CGG CTC CTT AGC ACC TCT 1792 Asn Val Ala Thr Leu Gly Leu Leu Met Ala Arg Leu Leu Ser Thr Ser 555 560 565 CCA GCT CTC CAA GGA ACC CCA GCC TCC CGA GGT TTC TTC GCA GCT GCC 1840 Pro Ala Leu Gln Gly Thr Pro Ala Ser Arg Gly Phe Phe Ala Ala Ala 570 575 580 ATC CTC TTT CTG TCA CAG TCC CAT GTG GCA CGA GCC ACC CCT GGC TCT 1888 Ile Leu Phe Leu Ser Gln Ser His Val Ala Arg Ala Thr Pro Gly Ser 585 590 595 GAC CAG GCA GTG TTG GCC CTG TCC CCT GAC TAT GAA GGC ATC TGG GCT 1936 Asp Gln Ala Val Leu Ala Leu Ser Pro Asp Tyr Glu Gly Ile Trp Ala 600 605 610 GAC TTG CAA GAG CTC TGG TTC CTG GGC ATG CAG GCC TTC ACG GGT TGT 1984 Asp Leu Gln Glu Leu Trp Phe Leu Gly M et Gln Ala Phe Thr Gly Cys 615 620 625 630 GTG CCG CTG CTG CCC TGG CTG GCC CCT GCC GCC CTG CGC TCC CGC TGG 2032 Val Pro Leu Leu Pro Trp Leu Ala Pro Ala Ala Leu Arg Ser Arg Trp 635 640 645 CCA CAG GAG CTG CTA CAA CTG CTA GGT AGT GTA AGC CCC AAC TCC GTC 2080 Pro Gln Glu Leu Leu Gln Leu Leu Gly Ser Val Ser Pro Asn Ser Val 650 655 660 AAG CCT GAG ATG GTG GCT GCC TAC TAC CAG GGC GTG CTG GTG GAA TTG GCA 2128 Lys Pro Glu Met Val Ala Ala Tyr Gln Gly Val Leu Val Glu Leu Ala 665 670 675 CGG GCT AAC CGG CTA TGC CGG GAG GCC ATG AGG CTG CAG GCG GGT GAA 2176 Arg Ala Asn Arg Leu Cys Arg Glu Ala Met Arg Leu Gln Ala Gly Glu 680 685 690 GAA ACG GCC AGC CAT TAC CGA ATG GCT GCT TTG GAG CAG TGC CTG TCA 2224 Glu Thr Ala Ser His Tyr Arg Met Ala Ala Leu Glu Gln Cys Leu Ser 695 700 705 710 GAG CCC TGAGGGGCAT CCAGTGGGAGGAGAGG Glu Pro GAGGAGGCAT CTTCCCTGAA G 2301

SEQ ID NO: 8 Sequence Length: 712 Sequence Type: Amino Acid Topology: Linear Sequence Type: Peptide Sequence Met Ala Ser Asp Cys Glu Pro Ala Leu Asn Gln Ala Glu Gly Arg Asn 5 10 15 Pro Thr Leu Glu Arg Tyr Leu Gly Ala Leu Arg Glu Ala Lys Asn Asp 20 25 30 Ser Glu Gln Phe Ala Ala Leu Leu Leu Val Thr Lys Ala Val Lys Ala 35 40 45 Gly Asp Ile Asp Ala Lys Thr Arg Arg Arg Ile Phe Asp Ala Val Gly 50 55 60 Phe Thr Phe Pro Asn Arg Leu Leu Thr Thr Lys Glu Ala Pro Asp Gly 65 70 75 80 Cys Pro Asp His Val Leu Arg Ala Leu Gly Val Ala Leu Leu Ala Cys 85 90 95 Phe Cys Ser Asp Pro Glu Leu Ala Ala His Pro Gln Val Leu Asn Lys 100 105 110 Ile Pro Ile Leu Ser Thr Phe Leu Thr Ala Arg Gly Asp Pro Asp Asp 115 120 125 Ala Ala Arg Arg Ser Met Ile Asp Asp Thr Tyr Gln Cys Leu Thr Ala 130 135 140 Val Ala Gly Thr Pro Arg Gly Pro Arg His Leu Ile Ala Gly Gly Thr 145 150 155 160 Val Ser Ala Leu Cys Gln Ala Tyr Leu Gly His Gly Tyr Gly Phe Asp 165 170 175 Gln Ala Leu Ala Leu Leu Val Gly Leu Leu Ala Ala Ala Glu Thr Gln 180 185 190 Cys Trp Lys Glu Ala Glu Pro Asp Leu Leu Ala Val Leu Arg Gly Leu 195 200 205 Ser Glu Asp Phe Gln Lys Ala Glu Asp Ala Ser Lys Phe Glu Leu Cys 210 215 220 Gln Leu Leu Pro Leu Phe Leu Pro Pro Thr Thr Val Pro Pro Glu Cys 225 230 235 240 His Arg Asp Leu Gln Ala Gly Leu Ala Arg Ile Leu Gly Ser Lys Leu 245 250 255 Ser Ser Trp Gln Arg Asn Pro Ala Leu Lys Leu Ala Ala Arg Leu Ala 260 265 270 270 His Ala Cys Gly Ser Asp Trp Ile Pro Val Gly Ser Ser Gly Ser Lys 275 280 285 Phe Leu Ala Leu Leu Val Asn Leu Ala Cys Val Glu Val Arg Leu Ala 290 295 300 Leu Glu Glu Thr Ser Thr Glu Val Lys Glu Asp Val Val Thr Ala Cys 305 310 315 320 Tyr Ala Leu Met Glu Leu Gly Ile Gln Glu Cys Thr Arg Cys Glu Gln 325 330 335 Ser Leu Leu Lys Glu Pro Gln Lys Val Gln Leu Val Ser Ile Met Lys 340 345 350 Glu Ala Ile Gly Ala Val Ile His Tyr Leu Leu Gln Val Gly Pro Glu 355 360 365 Lys Gln Lys Glu Pro Phe Val Phe Ala Ser Val Arg Ile Leu Gly Ala 370 375 380 380 Trp Leu Ala Glu Glu Thr Ser Ser Leu Arg Lys Glu Val Cys Gln Leu 385 390 395 400 Leu Pro Phe Leu Val Arg Tyr Ala Lys Thr Leu Tyr Gly Glu Ala Glu 405 410 415 Glu Ala Ser Asp Ile Ser Gln Gln Val Ala Asn Leu Ala Ile Ser Pro 420 425 430 Thr Thr Pro Gly Pro Ser Trp Pro Gly Asp Ala Leu Arg Leu Leu Leu 435 440 445 Pro Gly Trp Cys His Leu Thr Val Glu Asp Gly Pro Arg Glu Ile Leu 450 455 460 Ile Lys Glu Gly Ala Pro Ser Leu Leu Cys Lys Tyr Phe Leu Gln Gln 465 470 475 480 Trp Glu Leu Thr Ser Pro Gly His Asp Thr Ser Val Leu Pro Asp Ser 485 490 495 Val Glu Ile Gly Leu Gln Thr Cys Cys His Ile Phe Leu Asn Leu Val 500 505 510 Val Thr Ala Pro Gly Leu Ile Lys Arg Asp Ala Cys Phe Thr Ser Leu 515 520 525 Met Asn Thr Leu Met Thr Ser Leu Pro Ser Leu Val Gln Gln Gln Gly 530 535 535 540 Arg Leu Leu Leu Ala Ala Asn Val Ala Thr Leu Gly Leu Leu Met Ala 545 550 555 560 Arg Leu Leu Ser Thr Ser Pro Ala Leu Gln Gly Thr Pro Ala Ser Arg 565 570 575 Gly Phe Phe Ala Ala Ala Ile Leu Phe Leu Ser Gln Ser His Val Ala 580 585 590 Arg Ala Thr Pro Gl y Ser Asp Gln Ala Val Leu Ala Leu Ser Pro Asp 595 600 605 Tyr Glu Gly Ile Trp Ala Asp Leu Gln Glu Leu Trp Phe Leu Gly Met 610 615 620 Gln Ala Phe Thr Gly Cys Val Pro Leu Leu Pro Trp Leu Ala Pro Ala 625 630 635 640 Ala Leu Arg Ser Arg Trp Pro Gln Glu Leu Leu Gln Leu Leu Gly Ser 645 650 655 Val Ser Pro Asn Ser Val Lys Pro Glu Met Val Ala Ala Tyr Gln Gly 660 665 670 Val Leu Val Glu Leu Ala Arg Ala Asn Arg Leu Cys Arg Glu Ala Met 675 680 685 Arg Leu Gln Ala Gly Glu Glu Thr Ala Ser His Tyr Arg Met Ala Ala 690 695 700 Leu Glu Gln Cys Leu Ser Glu Pro 705 710

[Brief description of the drawings]

FIG. 1 shows C16N-1 in various cells and tissues.
It is an electrophoresis photograph as a result of having investigated the expression of / C16N-2 and C16N protein by Western blot analysis.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁶ Identification code FI A61K 38/00 AAM C07K 14/48 ABJ 16/18 ADD C12P 21/02 C C07K 14/47 A61K 37/02 AAB 14/48 AAM 16/18 ABJ C12N 5/10 ADD C12P 21/02 C12N 5/00 B // (C12N 5/10 C12R 1:91) (C12P 21/02 C12R 1:91)

Claims (11)

[Claims] 1. A DNA encoding a protein consisting of the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8. 2. The DNA according to claim 1, comprising the nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 7. 3. A DNA which hybridizes with the DNA according to claim 1 or 2 under stringent conditions and has the following properties (1), (2), (3) and / or (4): DNA encoding a non-secreted protein having the same. (1) It has an activity of inducing differentiation of bone marrow cells into cells having hydroxyapatite decomposability. (2) It has an activity to maintain the survival of nerve cells. (3) It has osteoblast proliferation inhibitory activity. (4) It has an activity of promoting the expression of type I collagen in osteoblasts. 4. An amino acid sequence shown in SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8 in which one or more amino acids have been deleted, substituted and / or added. A DNA encoding a non-secretory protein having the following properties (1), (2), (3) and / or (4): (1) It has an activity of inducing differentiation of bone marrow cells into cells having hydroxyapatite decomposability. (2) It has an activity to maintain the survival of nerve cells. (3) It has osteoblast proliferation inhibitory activity. (4) It has an activity of promoting the expression of type I collagen in osteoblasts. A protein obtained by expressing the DNA according to any one of claims 1 to 4. 6. An expression vector containing the DNA according to any one of claims 1 to 4. 7. A transformant transformed by the expression vector according to claim 6. 8. A method for producing a recombinant protein, comprising culturing the transformant according to claim 7, and recovering the expressed recombinant protein. 9. A medicament comprising the DNA according to claim 1 or the protein according to claim 5 as an active ingredient. 10. An antibody against the protein according to claim 5. 11. A transgenic animal in which the DNA according to any one of claims 1 to 4 is artificially introduced into a chromosome, or any of which is deleted from the chromosome.