KR102136643B1 - Novel hepatocellular carcinoma diagnostic marker and use thereof - Google Patents

Abstract

The present invention relates to a novel liver cancer diagnostic marker and uses thereof, and a novel biomarker composition for predicting and diagnosing liver cancer, a composition for diagnosing liver cancer, a liver cancer diagnostic kit, and a method for providing information for predicting and diagnosing liver cancer. The novel liver cancer diagnostic markers according to the present invention can be usefully used for the diagnosis of liver cancer, as it is confirmed that the expression level is increased in proliferative liver cancer cells caused by monounsaturated fatty acids.

Description

Novel hepatocellular carcinoma diagnostic marker and use thereof

The present invention relates to novel liver cancer diagnostic markers and uses thereof.

Hepatocellular carcinoma (HCC) is one of the most fatal cancers in the world. Particularly in Asia and sub-Saharan Africa each year has been a more than five hundred thousand people died of liver cancer, and if the country is also increasing year by year the number of liver cancer patients the occurrence of liver cancer is often endemic to the inactivation of tumor suppressor genes such as p53 and p16 ^INK4A And activated by other unknown carcinogens (oncogenes). In addition, hepatitis B and hepatitis C virus infections and exposure to aflatoxin β1 in liver tissue have been reported as causes, and several growth factors such as transforming growth factor α (TGF-α) and TGF-β factors) have been reported to be involved in the progression of liver cancer. In addition, there have been reports that DNA mutations and genetic alteration of gene expression are confirmed in patient tissues of liver cancer.

On the other hand, there is liver resection as a fundamental treatment for liver cancer, but only 30-40% of patients diagnosed early can perform surgical treatment. Conservative anticancer treatment for advanced liver cancer that cannot perform surgery has low effectiveness and high recurrence rate. Early diagnosis is very important.

As a method currently used for the diagnosis of liver cancer, ultrasound, CT, MRI, and other diagnostic imaging tests, serum tests (AFP, PIVKA-II), and pathological biopsy of biopsy materials are used.

In addition, recently, a diagnostic method using a tumor marker has been used to diagnose cancer early, and since tumor markers are often overexpressed in specific cancer cells, it is confirmed that tumor markers have been expressed above a standard value, and thus, cancer of a specific organ is confirmed. Because it can diagnose. In addition, the diagnosis of cancer using a tumor marker is mostly performed by a blood test method using antibodies, and some are also tested using tissue extract, urine, feces, and the like. Alpha-fetoprotein, that is, alpha-fetoprotein (AFP) is used as a representative tumor marker for liver cancer. AFP is a glycoprotein that is abnormally produced when liver cells are transformed and dedifferentiated into cancer cells. Primary liver cancer , Hepatitis, cirrhosis, and fetal cancer, and is used for observation of treatment progress.

However, in the case of the AFP liver cancer marker used in the related art, the diagnosis rate for this is only 40-50%, and there is a disadvantage that the difference between the patients with fatty liver and hepatitis is not improved. Therefore, there is an urgent need to develop a new tumor marker that can effectively increase the diagnosis rate of early liver cancer by reinforcing these drawbacks.

On the other hand, fatty acid (fatty acid) is reported to be increased in cancer showing a high proliferation rate, it is known to regulate the secreted protein from cancer cells. For example, linoleic acid is known to increase the secretion of plasminogen activator inhibitor-1 in breast cancer, and oleate increases MMP9 secretion in breast cancer cells. It is known to induce cancer invasion, and it is known to increase the likelihood of hepatitis by increasing the secretion of interleukin-8 from steroidal liver cancer cells. In particular, among the types of fatty acids, oleic acid is the most abundant circulating free FA in mammals, and it is known that the level of fatty acids in cancer tissues is increased.

However, studies on factors affecting monounsaturated fatty acid oleate on liver cancer and changes in expression in liver cancer cells or liver cancer tissues due to oleate fatty acids have been incomplete.

Republic of Korea Registered Patent No. 10-0325766 Republic of Korea Registered Patent No. 10-1351234

Accordingly, the present inventors discovered genes with increased expression levels in liver cancer cells, specifically, proliferative liver cancer cells caused by monounsaturated fatty acids, and confirmed that these genes can be used as new biomarkers for liver cancer diagnosis. Was completed.

Accordingly, an object of the present invention is to provide a novel biomarker composition for predicting and diagnosing liver cancer.

Another object of the present invention is to provide a novel composition for diagnosing liver cancer.

Another object of the present invention is to provide a liver cancer diagnostic kit comprising the composition for diagnosing liver cancer of the present invention.

Another object of the present invention is to provide a method for providing information for predicting and diagnosing liver cancer.

In order to achieve the above object, the present invention provides a biomarker composition for predicting and diagnosing liver cancer, comprising at least one gene selected from among the 18 genes listed in the table below or a protein expressed from the gene.

In one embodiment of the present invention, when the gene is treated with a monounsaturated fatty acid, the expression level may increase in liver cancer cells.

In one embodiment of the present invention, the biomarker composition may be to determine whether proliferation of liver cancer cells by monounsaturated fatty acids.

In addition, the present invention provides a composition for diagnosing liver cancer, comprising a substance for measuring mRNA or protein levels of one or more genes selected from among 18 genes listed in the table above.

In one embodiment of the present invention, the substance may be a primer, probe or antibody that specifically binds to one or more genes or proteins selected from 18 genes listed in the table.

In one embodiment of the present invention, the composition for diagnosing liver cancer may be to determine whether proliferation of liver cancer cells by monounsaturated fatty acids.

In addition, the present invention provides a liver cancer diagnostic kit comprising the composition for diagnosing liver cancer of the present invention.

Furthermore, the present invention, (a) measuring the mRNA or protein expression level of the 18 genes listed in the table of claim 1 from a biological sample isolated from the patient; And (b) measuring the amount of mRNA or protein expression of the gene from the normal control sample and comparing it with the measurement result of step (a), providing a method for providing information for predicting and diagnosing liver cancer.

In one embodiment of the present invention, the biological sample may be liver tissue, liver cells isolated from the subject, or a culture medium thereof.

In one embodiment of the present invention, if the mRNA or protein expression level of the gene is increased compared to the normal control, it may be further comprising the step of determining that the liver cancer has developed or is proliferative liver cancer.

In one embodiment of the present invention, the liver cancer may be a proliferative liver cancer having cancer cell proliferative activity by monounsaturated fatty acids.

The present invention relates to a novel biomarker composition for predicting and diagnosing liver cancer, a composition for diagnosing liver cancer, a kit for diagnosing liver cancer, and a method for providing information for predicting and diagnosing liver cancer. The novel liver cancer diagnosis markers according to the present invention can be usefully used for the diagnosis of liver cancer as these markers are found to increase when treatment with a monounsaturated fatty acid that causes liver cancer.

1 is a schematic diagram showing an experimental process for analyzing a change pattern of a secreted protein body according to oleate treatment for a liver cancer cell line (HepG2) in one embodiment of the present invention.
Figure 2 confirms the cell proliferation-promoting action of hepatocellular carcinoma cell line according to the oleate treatment, 2a is to confirm the proliferation of hepatic cancer cells according to the concentration of oleate treatment, 2b is a group treated with palmitate and oleate and oleate It was a comparative analysis of the degree of proliferation of liver cancer cells in the treatment group alone.
Figure 3 shows the chromatogram of the base peak according to the LC-MS/MS experiment in the group treated with oleate (Ole-CM#) and the group not treated with oleate (NT-CM#).
FIG. 4 shows the similarity for ID (4a) and intensity (4b) for 24 LC-MS/MS datasets obtained by LC-MS/MS analysis in the form of a heat map, oleated. The group was indicated as'Oleate treated', the oleate-untreated group was indicated as'Control', and the color bar indicates the degree of similarity score.
Figure 5 shows the number of peptides identified using MS-GF+ analysis results for the oleate-treated and oleate-untreated groups (FIG. 5A), and the number of peptides identified using AMT-DB (FIG. 5B).
FIG. 6 shows that the secreted proteins up-regulated by oleate treatment in the liver cancer cell line are analyzed and classified according to cell function.

The present invention is characterized by providing a novel biomarker and its use for predicting and diagnosing liver cancer.

The present inventors were studying to discover biomarkers capable of accurately and effectively predicting and diagnosing liver cancer, and showed genes showing differences in expression levels in the group treated with unsaturated fatty acids and the group not treated with liver cancer cell lines. Pity.

Specifically, among the types of monounsaturated fatty acids, it was confirmed through the following experiment that oleate induces proliferation of cancer cells, and genes whose expression is increased in liver cancer cells by oleate treatment were identified through profile analysis of secreted protein bodies. , Among these, it was confirmed that a total of 18 genes can be used as a novel biomarker for prediction and diagnosis of liver cancer.

Accordingly, the present invention can provide a biomarker composition for predicting and diagnosing liver cancer, comprising one or more genes selected from among the 18 genes listed in the following table or proteins expressed from the genes.

In addition, the 18 genes listed in the table can determine whether or not proliferation of hepatocellular carcinoma cells by oleate, that is, the expression of one or more genes selected from the 18 genes is increased compared to the normal control in the sample obtained from the patient. If present, it may be judged that there is a possibility of proliferation of liver cancer cells or a possibility of developing liver cancer.

Therefore, the present invention can provide a composition for diagnosing liver cancer, comprising a substance for measuring mRNA or protein levels for one or more genes selected from among 18 genes listed in the table above.

Further, the substance may be a primer, probe or antibody that specifically binds to one or more genes or proteins selected from the 18 genes listed in the table.

In the present invention, the biomarker is a polypeptide or nucleic acid (e.g., mRNA, etc.), lipids, glycolipids, glycoproteins, sugars (monosaccharides, disaccharides, oligosaccharides) that show an increase or decrease in expression in liver tissue or liver cells during the onset of liver cancer. Organic biomolecules, etc.). The biomarker provided by the present invention may be a protein in which the 18 genes or the genes expressed in the expression level are increased in liver cells treated with monounsaturated fatty acid, preferably oleate, compared to normal liver tissue.

In the present invention, the expression level of a gene, preferably means the mRNA level at which the gene is expressed, that is, the amount of mRNA, and a substance capable of measuring the level may include a primer or probe specific to the gene. have. In the present invention, a primer or probe specific for any one or more genes selected from among the 18 may be a primer or probe capable of specifically amplifying the entire gene or a specific region of the gene, and the primer or probe is a sugar. It can be designed through methods known in the industry.

The term primer in the present invention is a single-strand capable of acting as a starting point for template-directed DNA synthesis under suitable conditions (i.e., four different nucleoside triphosphates and polymerases) in a suitable temperature and a suitable buffer. Means oligonucleotide. The suitable length of the primer may vary depending on various factors, such as temperature and the use of the primer. In addition, the sequence of the primer does not need to have a sequence completely complementary to some of the sequence of the template, it is sufficient to have sufficient complementarity within a range capable of hybridizing with the template and working with the primer. Therefore, the primer in the present invention does not need to have a sequence perfectly complementary to the nucleotide sequence of the template gene, and it is sufficient to have sufficient complementarity within a range capable of hybridizing to this gene sequence and acting as a primer. In addition, it is preferable that the primer according to the present invention can be used for a gene amplification reaction.

The amplification reaction refers to a reaction for amplifying a nucleic acid molecule, and the amplification reactions of these genes are well known in the art, for example, polymerase chain reaction (PCR), reverse transcriptase chain reaction (RT-PCR), and Riga Aze chain reaction (LCR), electron-mediated amplification (TMA), nucleic acid sequence substrate amplification (NASBA), and the like may be included.

In the present invention, the term probe refers to a linear oligomer of natural or modified monomers or linkages, includes deoxyribonucleotides and ribonucleotides and can specifically hybridize to the target nucleotide sequence, and is naturally present. It means synthetically or artificially. The probe according to the present invention may be a single chain, preferably an oligodioxyribonucleotide. Probes of the present invention may include natural dNMPs (ie, dAMP, dGMP, dCMP and dTMP), nucleotide analogues or derivatives. In addition, the probe of the present invention may also include ribonucleotides. For example, the probe of the present invention may be a skeleton modified nucleotide, such as peptide nucleic acid (PNA) (M. Egholm et al., Nature, 365:566-568 (1993)), phosphorothioate DNA, phosphorodithioate DNA, phosphoramidate DNA, amide-linked DNA, MMI-linked DNA, 2'-O-methyl RNA, alpha-DNA and methyl phosphonate DNA, sugar modified nucleotides such as 2'-O-methyl RNA, 2'-fluoro RNA, 2'-amino RNA, 2'-O-alkyl DNA, 2'-O-allyl DNA, 2'-O-alkynyl DNA, hexose DNA, pyranosyl RNA and anhydrohexyl Tall DNA, and nucleotides with base modifications, such as C-5 substituted pyrimidines (substituents are fluoro-, bromo-, chloro-, iodo-, methyl-, ethyl-, vinyl-, formyl-, e Tityl-, propynyl-, alkynyl-, thiazoryl-, imidazoryl-, pyridyl-), 7-deazapurine with C-7 substituents (substituents are fluoro-, bromo-, chloro- , Iodo-, methyl-, ethyl-, vinyl-, formyl-, alkynyl-, alkenyl-, thiazolyl-, imidazoryl-, pyridyl-), inosine and diaminopurine .

In addition, as a substance capable of measuring the level of the protein in the present invention, antibodies such as polyclonal antibodies, monoclonal antibodies, and recombinant antibodies capable of specifically binding to the protein expressed from the marker gene of the present invention can be used. Can contain

The “antibody” may be prepared using a technique known by a person skilled in the art. For the preparation of the antibody, for example, in the case of a polyclonal antibody, the antigen of the protein is injected into an animal and blood is collected from the animal. Can be produced by methods well known in the art to obtain serum containing the antibody, and these polyclonal antibodies can be obtained from any animal species host such as goat, rabbit, sheep, monkey, horse, pig, cow, dog, etc. It can be manufactured. In the case of monoclonal antibodies, it can be prepared using a hybridoma method well known in the art (Kohler et al., European Jounral of Immunology, 6, 511-519, 1976) or a phage antibody library ( Clackson et al, Nature, 352, 624-628, 1991, Marks et al, J. Mol. Biol., 222:58, 1-597, 1991). In addition, the antibody according to the present invention may include functional fragments of antibody molecules, as well as complete forms having two full-length light chains and two full-length heavy chains. A functional fragment of an antibody molecule means a fragment having at least an antigen-binding function, and includes Fab, F(ab'), F(ab') 2, and Fv. In addition, the present invention may provide a liver cancer diagnostic kit comprising the marker for diagnosing liver cancer according to the present invention or the composition for diagnosing liver cancer.

The liver cancer diagnostic kit of the present invention may include primers, probes or antibodies capable of measuring the expression level of the marker gene or the amount of protein expressed from the gene, and their definitions are as described above.

If the liver cancer diagnostic kit of the present invention is applied to a PCR amplification process, the kit of the present invention is optionally a reagent required for PCR amplification, such as a buffer solution, a DNA polymerase (eg, Thermus aquaticus (Taq), Thermus thermophilus (Tth) ), Thermus filiformis, Thermis flavus, Thermococcus literalis or thermally stable DNA polymerase obtained from Pyrococcus furiosus (Pfu)), DNA polymerase cofactors and dNTPs, and the liver cancer diagnostic kit of the present invention is applied to immunoassays. In some cases, the kit of the present invention may optionally include a secondary antibody and a substrate of a label.

Furthermore, the kit according to the present invention can be manufactured in a number of separate packaging or compartments containing the above-described reagent components.

In addition, the present invention can provide a microarray for diagnosing liver cancer comprising the biomarker for diagnosing liver cancer.

In the microarray of the present invention, a primer, probe, or antibody capable of measuring the expression level of the marker protein or a gene encoding the marker protein is used as a hybridizable array element and immobilized on a substrate. . Preferred substrates are suitable rigid or semi-rigid supports, such as membranes, filters, chips, slides, wafers, fibers, magnetic beads or non-magnetic beads, gels, tubing, plates, polymers, microparticles and capillaries. have. The hybridization array elements are arranged and immobilized on the substrate, and such immobilization can be performed by a chemical bonding method or a covalent bonding method such as UV. For example, the hybridization array element can be bonded to a glass surface modified to include an epoxy compound or an aldehyde group, and can also be bonded by UV at a polylysine coated surface. In addition, the hybridizing array elements can be bound to the substrate via linkers (eg, ethylene glycol oligomers and diamines).

Meanwhile, when the sample applied to the microarray of the present invention is a nucleic acid, it can be labeled and hybridized with an array element on the microarray. Hybridization conditions may be varied, and detection and analysis of the degree of hybridization may be variously performed depending on the labeling substance.

Furthermore, the present invention can provide a method for providing information for predicting and diagnosing liver cancer. Preferably, the method comprises: (a) mRNA or protein expression of 18 genes listed in the table of claim 1 from biological samples isolated from the patient. Measuring; And (b) measuring the mRNA or protein expression level of the gene from the normal control sample and comparing it with the measurement result of step (a).

The method of measuring the expression level of the gene or the amount of protein in the above may be performed by including a known process for separating mRNA or protein from a biological sample using a known technique.

In the present invention, the "biological sample" refers to a sample taken from a living body whose expression level or protein level is different from a normal control according to the degree of occurrence or progression of liver cancer, and the sample is, for example, Although not limited, tissue, cells, blood, serum, plasma, saliva and urine may be included, and preferably liver cancer tissue, liver cancer cells, or a culture medium thereof.

Measurement of the expression level of the gene is preferably to measure the level of mRNA, and the method of measuring the level of mRNA is reverse transcriptase chain reaction (RT-PCR), real-time reverse transcriptase chain reaction, RNase protection assay, Northern Blots and DNA chips, but are not limited thereto.

In addition, the method for measuring the amount of protein or the activity of the protein may be performed through various methods known in the art, for example, but not limited to, Western blot, Northern blot, ELISA (enzyme linked immunosorbent assay) ), radioimmunoassay (RIA), radioimmunodiffusion and immunoprecipitation assay.

For the measurement of the protein level, an antibody may be used. In this case, the marker protein in the biological sample and the antibody specific to this form a binding agent, that is, an antigen-antibody complex, and the amount of formation of the antigen-antibody complex is a detection label. It can be quantitatively measured through the signal level of (detection label). The detection label may be selected from the group consisting of enzymes, fluorescent substances, ligands, luminescent substances, microparticles, redox molecules, and radioactive isotopes, but is not limited thereto. As an analytical method for measuring protein levels, but is not limited to, Western blot, ELISA, radioimmunoassay, radioimmunoassay, Oukteroni immunodiffusion method, rocket immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, Complement fixation, FACS, and protein chips.

Therefore, the present invention can confirm the amount of mRNA expression or protein amount of the marker gene in the liver cancer patient or suspected liver cancer patient and the amount of protein expression of the marker gene of the control group through the above detection methods, the amount of expression By comparing the degree of control with the control group, it is possible to predict and diagnose the onset, progression or prognosis of liver cancer.

More specifically, the method for predicting or diagnosing the onset of liver cancer is determined to be caused by liver cancer or proliferative liver cancer when the expression level of the marker gene or the amount of the expressed protein according to the present invention is increased compared to the normal control sample. Can.

In addition, the proliferative liver cancer may be a proliferative liver cancer in which proliferation of cancer cells is promoted by a monounsaturated fatty acid, preferably the monounsaturated fatty acid may be oleate.

As described above, the 18 novel liver cancer biomarkers for diagnosis and prediction identified in the present invention are secreted proteins with increased expression and increased secretion in liver cancer cells having cancer cell proliferation activity by monounsaturated fatty acids, By measuring the expression level, liver cancer can be easily predicted and diagnosed.

Hereinafter, the present invention will be described in more detail by examples. These examples are only for explaining the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited to these examples.

< Preparation example And experiment method>

Reagent and cell culture

Sodium oleate (O7501, St. Louis, MO) and sodium palmitate (P9767, St. Louis, MO) were purchased from Sigma Aldrich. Fetal bovine serum (BSA), fraction V and fatty acid-free (126575, San Diego, CA) were purchased from Calbiochem and used. Olate or palmitate was used by dissolving in 100% methanol, and free fatty acids were combined with BSA, where fatty acid to BSA was used in a molar ratio of 5:1. In addition, when these cells were treated with cells, they were diluted with serum-free MIN (Minimum Essential Medium) and used to prepare a suitable final concentration roll. HepG2 cells were cultured using MEM (Welgene, LM 007-07) medium, and then 10% (v/v) of FBS (Lonza), 2 mM glutamine and 1% penicillin-streptomycin (Gibco) were added and 37°C was added. , 5% CO2 and 95% humidity conditions.

Cell proliferation Assay

Cell proliferation was assayed for cell viability using MTT (Sigma Aldrich, M5655) as a colorimetric assay. After dispensing HepG2 cells in a 96-well plate and incubating for 24 hours, the medium was incubated for 12 hours without addition, and then cultured by adding fatty acids for 24 hours, followed by secretome analysis. Samples were prepared. At this time, the control sample was used instead of the fatty acid treated with methanol and BSA of the same concentration. Cells were treated with 0.5 mg/ml MTT reagent, diluted with phenol-red medium, and cultured in a 37° C. CO ₂ incubator. After 2 hours, 100 μl DMSO was added to dissolve the formazan crystal. Intensity measurements were taken using a microplate reader at 540 nm absorbance.

Sample preparation

HepG2 cells were kept starved for 12 hours before fatty acid treatment, and then washed three times with 1xPBS to remove secretory factors secreted from these cells. For preparation of the sample treated with oleic acid, BSA-bound oleic acid (Oleate) was diluted with serum-free medium to a final concentration of 500 uM, treated, and incubated for 24 hours. As a control, a serum-free medium without oleic acid was treated with the same amount. In addition, the conditioned media was prepared by replacing the unrepleted serum-free medium with incubation in a 37° C. CO ₂ incubator for 24 hours. Thereafter, the culture medium was centrifuged at 3000 rpm for 10 minutes at 4° C., and the supernatant was filtered to remove contaminants, and the finally obtained conditioned media was lyophilized. The sample was prepared by drying.

Protein extraction and degradation

The dried sample obtained by the above method was dissolved with a lysis buffer (urea, NaCl, 50 mM Tris-HCl (pH 8.2)) and treated with a complex mini protease inhibitor tablet (Roche Applied Science, Basel, Switzerland). Thereafter, the ultracentrifuge was centrifuged at 45,000xg for 1 hour at 4°C. Thereafter, the protein concentration contained in the supernatant was quantified using a DC protein assay (Bio-Rad, Hercules, USA), and the protein sample was subjected to a reduction reaction at 37°C for 50 minutes using 0.5M DTT (100 μl). . Thereafter, 1.5 μl of 1M iodoacetamide was treated and an alkylation reaction was performed at room temperature for 30 minutes in dark conditions. After the reaction was completed, the sample was decomposed with trypsin (1:50 enzyme-to-protein ratio (w/w), Promega, Madison, WI, USA) and reacted overnight at 37°C. After completion of the reaction, the peptide samples were centrifuged at 2500xg for 10 minutes at room temperature, and the obtained aqueous solution was desalted through solid-phase extraction using reverse-phase tC18 Sep-Pak solid-phase extraction cartridges. Thereafter, the final peptide was eluted with a solvent of 50% ACN and 0.5% HAcO, dried in a spibbag and dissolved in a 0.1% formic acid solution. Thereafter, the sample was stored at 20°C until LC-MS/MS analysis.

LC-MS/MS analysis

All peptide samples are equipped with Thermo EASY-nLC 1000 (Thermo Scientific, Odense, Denmark) equipment equipped with an analytical column (Thermo Scientific, Easy-Column, 75 μm×50 cm) and a trap column (75 μm×2 cm). And separated. At this time, the operating temperature for the analytical column was set to 50° C., and the flow rate was set to 300 nl/min. As a solvent, water containing 0.1% formic acid and 2% acetonitrile was used, and solvent B was 0.1%. An acetonitrile solvent with formic acid and 2% water was used. For analysis of the desired proteome, peptides were analyzed under a concentration gradient condition for 120 minutes, and the concentration gradient was eluted with 2-40% solvent B for 90 minutes, and 40-80% with solvent B for 10 minutes, 80%. Elution was performed with solvent B for 10 minutes, and with elution of 80% to 2% solvent B for 10 minutes. The peptides separated from the LC were analyzed using a QExactive™ hybrid quadrupole-Orbitrap mass spectrometer (Thermo Scientific) equipped with a nanoelectrospray source, and the electrical potential of transcriptional injection ionization was 1.7 kV for ionization of the eluted peptide, The temperature of the desolvation capillary was set to 270°C. In addition, the Q-Exactive mass spectrometer was operated in data dependent mode to obtain irradiation scans for a mass range of 450-2000 Thomsons (Th) with a resolution of 70,000 (m/z 200). Then, for MS/MS analysis, the top 10 ions most contained in the irradiation scan were selected using a separation window of 2.0Th, and a high energy collision dissociation (HCD) method set to 25 standard collision energy and an exclusion period of 10 seconds Peptide ions were fragmented.

Subsequently, MS/MS scans were acquired at a resolution of 17,500, and the maximum ion implantation times for the entire MS and MS/MS scans were 100 ms and 50 ms, respectively. The automatic gain control (AGC) target values were set to 1.0×10 ⁶ and 1.0×10 ⁵ for each of the entire MS and MS/MS scans.

Peptide Sympathy

The spectrum of each fragment was generated as a mzXML file using MSConvert (ProteoWizard release: 3.0.4323) method. The MS data was firstly calibrated using PE-MMR (postexperiment-monoisotopic mass refinement) to assign the correct precursor mass to tandem MS data. It also uses the MS-GF + Beta (v10089) analysis engine to target-decoy for a human-reference proteome (UniProtKB release 2015_11, a database of UniProtKB / Swiss-Prot of 42,123 proteins and a composite database containing a list of 179 common contaminating proteins) MS/MS spectra were searched by setting, and the following parameters were analyzed: semitryptic, precursor mass tolerance 10 ppm, carbamidomethylation of cysteine as a fixed modification option, and methionine oxidation as a variable modification option. In addition, all the search results of 24 LC-MS/MS data sets were combined, and a target-decoy analysis was applied to the combined data set to identify peptides having a false detection rate (FDR) of 0.01 or less.

Uncovered Peptide Quantitative analysis

In order to assign MS intensity to each of the identified peptides, a non-labeling method based on MS intensity was applied to 24 LC-MS/MS data sets as previously described. Briefly, MS spectra of peptides that appeared consecutively at a specific time during LC elution in LC-MS/MS experiments through PE-MMR analysis were classified as intrinsic mass grade (UMC). Ideally, each UMC corresponds to one peptide, and includes mass spectral features such as the ions corresponding to the peptides and their charge state, abundance (intensity), scan number and measured single isotope mass. For each UMC, the UMC mass was calculated by calculating the intensity-weighted average of the monoisotopic mass of all ions belonging to the UMC. For peptides corresponding to UMC, the abundance of the corresponding peptide was evaluated as the sum of the abundance of all ions of UMC (UMC intensity). In addition, the precursor mass of the MS/MS spectrum corrected by PE-MMR was matched with the UMC mass, and the corresponding value was replaced with the mass of the UMC. In this process, MS/MS spectrum information was linked to the corresponding UMC. The linked MS/MS spectrum was assigned to a peptide sequence (peptide ID) of FDR <0.01 calculated by previous MS-GF + search and target-decoy analysis. The peptide ID was assigned to the UMC, and the UMC intensity was assigned to the intensity value of the peptide ID.

Based on master precision mass/time tag database UMC's Assignment

To assign peptide IDs to unidentified UMC, a master precision mass/time tag (AMT) database (DB) was prepared and used as previously described. Briefly, information on UMC having a peptide ID of 24 LC-MS/MS data (LC-MS/MS analysis of 8 samples repeated 3 times) was collected in the form of a master AMT DB. AMT is a unique peptide sequence determined by experimentally determined single isotopic masses and standardized elution time (NET), and the average mass of the peptide and the median of NET when the peptide corresponding to each AMT is measured multiple times Was recorded in AMT. Thereafter, an unidentified UMC was mapped to the AMT of the master AMT DB by allowing a mass of ± 10 p.p.m. and a NET error of ± 0.02. It was confirmed that UMCs not identified in the mapping process were mapped to AMTs within mass and NET tolerances, and Xcorr measurement values were calculated to evaluate the similarity between MS/MS spectra of AMTs and UMCs. Finally, the unidentified UMC was determined to match AMT when Xcorr was greater than 3.0, and for each unidentified UMC matching AMT, a peptide ID for AMT was assigned to the unidentified UMC, where AMT (UMC mass and NET All information for) was allocated together.

Sympathetic Peptide Sorting process

After allocating the unidentified UMC using the master AMT DB, the allocation of the LC-MS/MS data set and the identified UMC were organized into nm sorting tables (peptide IDs for n UMC and UMC intensity shown in m samples). . Missing values in the alignment table indicate that the peptide was not identified in the corresponding data set. To fill in the missing values in each row of the table, additional unrecognized UMCs that were likely to match UMCs sorted by UMC mass and NET were searched with the following error tolerance criteria: ± for repeated datasets for the same sample. 10 ppm Tolerance of mass error and ±0.01 NET error, ±10 p.p.m. for other experimental samples. Tolerance of mass error and ±0.02 NET error. After the missing values of the sorting table were added by additional search, the normalization of the peptides was performed on the UMC intensity shown in the table to correct the amount of peptide present throughout the data set. In addition, for evaluating the reproducibility of LC-MS/MS analysis, the number of peptide IDs and peptide strengths overlapped between the LC-MS/MS data sets were measured, and each of the two types of similarity scores (ID similarity, intensity similarity) was measured. It was calculated.

Identification of proteins whose secretion amount changes significantly

In order to identify proteins with a significantly different secretion amount depending on the presence or absence of oleate treatment, the previously reported integrated statistical method was applied to the intensity of UMC (peptide) in the sorting table to significantly change the expression (DEpeptides) ) Was selected. Briefly explaining the statistical method, the log ₂ -intensity of each peptide in the oleated sample is determined by the Student's t-test and the T-value for each peptide and the mid-latitude test resulting in the log ₂ -median ratio. Compared to the log ₂ -intensity of the untreated control. Then, 24 samples were randomly repeated 1000 times to estimate the empirical null distribution of the T value and the log ₂ -intermediate ratio. Adjusted P values of the observed values of T and log ₂ -median ratio for each peptide were calculated using empirical distribution through two-sided tests, and integrated P values were calculated using Stouffer's method. DEpeptide was defined by the following criteria. First, when the corresponding peptide is detected in at least six samples in one of the conditions with or without oleate treatment, second, the combined P value of the peptide is less than 0.05 and the log ₂ -absolute value of the ratio of the peptide is log ₂ -In the case of the 95th percentile or more on the experimental distribution of the median ratio, the peptide was identified as DEpeptide. Further, in addition to the obtained DEpeptide, peptides that were not detected in all samples under one condition among the two conditions with or without treatment of oleate were selected as DEpeptides under other conditions. Lastly, for proteins with two or more native DEpeptide numbers that were increased or decreased during oleate treatment, they were identified by increasing or decreasing DSP, respectively.

Protein detected or DSPs Functional analysis

Analysis of Gene Ontology biological processes (GOBPs) or Genes and Genomes (KEGG) pathways from Kyoto Encyclopedia for protein lists of detected proteins or DSPs was analyzed using DAVID software. GOBP representing a specific protein list was identified as GOBP with P <0.01, and the P value of cell function was converted to Z-score using Z = -N ^-1 (P) for visualization, where N ^-1 is inverse Standard normal distribution.

Western Blotting

For Western blotting, HepG2 cells were collected using a lysis buffer (10 mM Tris-HCl, pH 7.4, 1.5 mM EDTA, 10% Glycerol) containing a protease cocktail (Roche Diagnostics, Indianapolis, USA). Subsequently, sonication and centrifugation were performed and proteins were quantified with Bradford reagent. The secretory (Secretome) sample was dissolved in a lysis buffer, concentrated by lyophilization, and mixed with SDS buffer. After the concentration gradient SDS-PAGE gel electrophoresis of 6 to 16% and transferred to a nitrocellulose membrane, blocking was performed with a TBS-T solution containing 5% skim milk for 30 minutes, and then 4°C as a primary antibody. Was reacted overnight, and then reacted with horseradish peroxidase bound anti-mouse or anti-rabbit antibody for 1 hour. Then, the membrane was washed with TBS-T solution and developed using ECL solution to analyze the protein expression level.

Data use

Mass spectrometry proteomics data were used deposited with the ProteomeXchange consortium through PRIDE (data set identifiers PXD007906 and 10.6019 / PXD007906).

< Example 1>

Oleate Confirmation of cell proliferation of liver cancer cells by fatty acid treatment

The present inventors first analyzed whether oleate fatty acids act to promote cell proliferation of liver cancer cells, and then treated with oleate to HepG2 cells, a liver cancer cell line for each concentration, and then analyzed whether or not proliferation of liver cancer cells through MTT analysis. Did.

As a result, it was found that HepG2 cell group treated with oleate had higher cell proliferation than cell group without oleate treatment. Therefore, it was found from these results that the fatty acid oleate can promote cell proliferation of liver cancer cell lines (see FIG. 2A ).

On the other hand, one of the saturated fatty acids, palmitate (palmitate) is known to have the activity of inhibiting the proliferation of cancer cells, the inventors of the oleate alone treatment group and oleate and palmitate to reconfirm the cancer cell proliferation activity The degree of cell proliferation in the Tate complex treatment group was analyzed.

As a result, as shown in FIG. 2B, it was found that the oleate-only treatment group had a significantly higher cancer cell proliferation than the oleate-palmitate combination treatment group.

As a result, it was found that oleate, one of the fatty acids, has an activity of promoting cell proliferation of liver cancer cells, and that liver cancer can develop and progress by cancer cell proliferation by oleate.

< Example 2>

Oleate Shows change in expression level by fatty acid treatment In liver cancer cell lines Secreted proteomic analysis

Furthermore, based on the results of Example 1, the present inventors performed mass spectrometry experiments as a method for discovering biomarkers capable of confirming whether liver cancer is caused by oleate fatty acids and proliferation of liver cancer cells. Among the proteins secreted from the cells, the secreted protein bodies showing a change in the expression level were screened for the group treated with oleic fatty acid and the group not treated with hepatic cancer cell line HepG2 cells. This analysis method was performed by the experimental method described above, and the schematic diagram of the experiment is shown in FIG. 1. Also, the cell lines of the oleate-treated group (n=4) and the untreated group (n=4) were subjected to repeated experiments three times, respectively, and the results were extracted.

<2-1> LC-MS/MS analysis results

After obtaining the culture medium of the oleic fatty acid-treated HepG2 cell group and the oleate-untreated HepG2 cell group, the proteins were hydrolyzed by trypsin treatment, and LC-MS/MS analysis was performed.

As a result, as shown in FIG. 3, 12 LC-MS/MS datasets were collected for the oleate-treated HepG2 cell population, and 12 LC-MS/MS datasets were collected for the oleate-untreated HepG2 cell population. A total of 24 LC-MS/MS datasets were obtained.

<2-2> Peptide Sympathy

For the 24 LC-MS/MS datasets obtained by the above method, PE-MMR analysis was performed using the MS-GF+ search method using the target decoy method to analyze the properties of the peptides, and UMC (confirmed from the 24 datasets) A master AMT DB containing 30,933 peptides was constructed using a unique mass class. Thereafter, AMT DB was used to match the peptides of the DB using mass and NET tolerances for unidentified UMC of individual samples, and then IDs were assigned to the matched peptides. After AMT DB search, peptides were quantified by UMC intensity in 24 LC-MS/MS data sets, and then arranged in a sort table. The reproducibility of unlabeled LC-MS/MS analysis was investigated using ID and intensity similarity scores by Mueller et al. using peptide abundances (UMC concentrations) normalized by Quantile normalization and 24 LCs. The mean ID and intensity similarity scores of the -MS/MS data set were 0.94 and 0.90, respectively, confirming high reproducibility of unlabeled LC-MS/MS analysis (see FIG. 4). In addition, based on 30,933 peptides in the AMT DB, 2640 HepG2 secreted proteins (2630 protein coding genes) with high reliability were identified as proteins with two or more unique peptides. The results of MS-GF+ analysis and AMT DB analysis according to the above method are shown in FIG. 5.

<2-3> Oleate Identification of secreted protein showing expression change according to treatment

The present inventors analyzed proteins showing an expression change among secreted proteins of liver cancer cells during oleate treatment. To this end, 30,933 peptides quantified in 24 LC-MS/MS sets (12 samples treated with oleate and 12 samples not treated with oleate) were constructed as a sorting table, and those in the oleated and untreated groups. Peptide abundances were compared and analyzed. That is, 2461 unique peptides (DEpeptide) were significantly increased or decreased by oleate treatment using the integrated statistical method described above for the aligned peptides (P <0.05).

As a result, when two unique peptides consistently showed an increase or decrease pattern by oleate treatment, this was defined as a differentially secreted protein (DSP), and as a result, 349 DSPs were identified. Of the 12 DSPs, two or more increased and decreased DEpeptide appeared simultaneously in one protein, thus excluding the DSP. Among the remaining DSPs, 145 were found to increase during oleate treatment, and 192 were found to decrease. Here, a list of 145 proteins increased in the liver cancer cell line by oleate is shown in Table 1 below.

145 proteins upregulated by oleate number UniProtKB accession Entrez ID Symbol Description Number of unique DEpeptides One Q96DT5 8701 DNAH11 dynein, axonemal, heavy chain 11 2 2 Q07021 708 C1QBP complement component 1, q subcomponent binding protein 2 3 P14174 4282 MIF macrophage migration inhibitory factor (glycosylation-inhibiting factor) 3 4 P06727 - APOA4 - 18 5 P13645 3858 KRT10 keratin 10, type I 3 6 P35637 2521 FUS FUS RNA binding protein 3 7 P52815 6182 MRPL12 mitochondrial ribosomal protein L12 3 8 P00505 2806 GOT2 glutamic-oxaloacetic transaminase 2 2 9 Q9BPW8 8508 NIPSNAP1 nipsnap homolog 1 (C. elegans) 2 10 Q92608 1794 DOCK2 dedicator of cytokinesis 2 2 11 P23588 1975 EIF4B eukaryotic translation initiation factor 4B 2 12 O15067 5198 PFAS phosphoribosylformylglycinamidine synthase 2 13 P02787 7018 TF transferrin 12 14 P61916 10577 NPC2 Niemann-Pick disease, type C2 2 15 P07306 432 ASGR1 asialoglycoprotein receptor 1 2 16 Q86VP6 55832 CAND1 cullin-associated and neddylation-dissociated 1 4 17 P49902 22978 NT5C2 5'-nucleotidase, cytosolic II 2 18 P10619 5476 CTSA cathepsin A 4 19 P53999 10923 SUB1 SUB1 homolog, transcriptional regulator 2 20 Q6IN85 55671 PPP4R3A protein phosphatase 4 regulatory subunit 3A 2 21 P02753 5950 RBP4 retinol binding protein 4 3 22 Q9NTK5 29789 OLA1 Obg-like ATPase 1 3 23 P06681 717 C2 complement component 2 2 24 O43852 813 CALU calumenin 7 25 P30566 158 ADSL adenylosuccinate lyase 3 26 Q15582 7045 TGFBI transforming growth factor beta induced 2 27 Q8NI22 90411 MCFD2 multiple coagulation factor deficiency 2 2 28 Q9H4A6 64083 GOLPH3 golgi phosphoprotein 3 3 29 Q6PCB0 64856 VWA1 von Willebrand factor A domain containing 1 5 30 Q8NBS9 81567 TXNDC5 thioredoxin domain containing 5 (endoplasmic reticulum) 3 31 O43847 4898 NRDC nardilysin convertase 2 32 O00410 3843 IPO5 importin 5 2 33 P78509 5649 RELN reelin 18 34 Q86V81 10189 ALYREF Aly/REF export factor 4 35 P07858 1508 CTSB cathepsin B 8 36 P08697 5345 SERPINF2 serpin peptidase inhibitor, clade F (alpha-2 antiplasmin, pigment epithelium derived factor), member 2 2 37 P27797 811 CALR calreticulin 4 38 P31937 11112 HIBADH 3-hydroxyisobutyrate dehydrogenase 3 39 P05067 351 APP amyloid beta precursor protein 5 40 P29622 5267 SERPINA4 serpin peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member 4 2 41 P51659 3295 HSD17B4 hydroxysteroid (17-beta) dehydrogenase 4 4 42 P18859 522 ATP5J ATP synthase, H+ transporting, mitochondrial Fo complex subunit F6 2 43 P02751 2335 FN1 fibronectin 1 4 44 Q15084 10130 PDIA6 protein disulfide isomerase family A member 6 6 45 Q92820 8836 GGH gamma-glutamyl hydrolase 4 46 P36955 5176 SERPINF1 serpin peptidase inhibitor, clade F (alpha-2 antiplasmin, pigment epithelium derived factor), member 1 5 47 Q14651 5357 PLS1 plastin 1 2 48 P62805 8294 HIST1H4I histone cluster 1, H4i 3 49 O60832 1736 DKC1 dyskeratosis congenita 1, dyskerin 2 50 P01019 183 AGT angiotensinogen 10 51 O95445 55937 APOM apolipoprotein M 5 52 P07602 5660 PSAP prosaposin 7 53 P04040 847 CAT catalase 3 54 P07686 3074 HEXB hexosaminidase subunit beta 4 55 P10909 1191 CLU clusterin 2 56 P20142 5225 PGC progastricsin (pepsinogen C) 2 57 Q14697 23193 GANAB glucosidase, alpha; neutral AB 3 58 P53634 1075 CTSC cathepsin C 2 59 P07237 5034 P4HB prolyl 4-hydroxylase subunit beta 5 60 P00751 629 CFB complement factor B 2 61 P10451 6696 SPP1 secreted phosphoprotein 1 3 62 Q6P988 147111 NOTUM notum pectinacetylesterase homolog (Drosophila) 5 63 P61626 4069 LYZ lysozyme 4 64 P02647 335 APOA1 apolipoprotein A-I 6 65 Q9H993 79624 ARMT1 acidic residue methyltransferase 1 2 66 O43405 1690 COCH cochlin 4 67 P01011 12 SERPINA3 serpin peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member 3 10 68 Q13162 10549 PRDX4 peroxiredoxin 4 3 69 P04114 338 APOB apolipoprotein B 118 70 P00450 1356 CP ceruloplasmin (ferroxidase) 16 71 P59998 10093 ARPC4 actin related protein 2/3 complex subunit 4 2 72 P26641 1937 EEF1G eukaryotic translation elongation factor 1 gamma 2 73 P25705 498 ATP5A1 ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit 1, cardiac muscle 2 74 P13473 3920 LAMP2 lysosomal associated membrane protein 2 2 75 P04066 2517 FUCA1 fucosidase, alpha-L-1, tissue 4 76 P02679 2266 FGG fibrinogen gamma chain 4 77 Q9UHG2 27344 PCSK1N proprotein convertase subtilisin/kexin type 1 inhibitor 3 78 P07339 1509 CTSD cathepsin D 3 79 P19022 1000 CDH2 cadherin 2 3 80 Q9H3G5 54504 CPVL carboxypeptidase, vitellogenic like 6 81 P06396 2934 GSN gelsolin 4 82 Q9BZZ5 8539 API5 apoptosis inhibitor 5 2 83 P14625 7184 HSP90B1 heat shock protein 90kDa beta family member 1 3 84 Q15181 5464 PPA1 pyrophosphatase (inorganic) 1 2 85 O15232 4148 MATN3 matrilin 3 4 86 P30101 2923 PDIA3 protein disulfide isomerase family A member 3 9 87 P50895 4059 BCAM basal cell adhesion molecule (Lutheran blood group) 2 88 Q9NYU2 56886 UGGT1 UDP-glucose glycoprotein glucosyltransferase 1 6 89 O95433 10598 AHSA1 AHA1, activator of heat shock 90kDa protein ATPase homolog 1 (yeast) 2 90 P02795 4502 MT2A metallothionein 2A 2 91 P18206 7414 VCL vinculin 2 92 O00584 8635 RNASET2 ribonuclease T2 2 93 Q14696 23184 MESDC2 mesoderm development candidate 2 2 94 P06865 3073 HEXA hexosaminidase subunit alpha 4 95 Q9BSE5 79814 AGMAT agmatinase 2 96 Q08830 2267 FGL1 fibrinogen like 1 4 97 P22792 1370 CPN2 carboxypeptidase N subunit 2 2 98 P28799 2896 GRN granulin 2 99 P05154 5104 SERPINA5 serpin peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member 5 4 100 P19823 3698 ITIH2 inter-alpha-trypsin inhibitor heavy chain 2 11 101 P14314 5589 PRKCSH protein kinase C substrate 80K-H 6 102 P31040 6389 SDHA succinate dehydrogenase complex flavoprotein subunit A 3 103 P42765 10449 ACAA2 acetyl-CoA acyltransferase 2 2 104 P13688 634 CEACAM1 carcinoembryonic antigen related cell adhesion molecule 1 2 105 Q02818 4924 NUCB1 nucleobindin 1 3 106 O60568 8985 PLOD3 procollagen-lysine, 2-oxoglutarate 5-dioxygenase 3 3 107 Q9Y2B0 10330 CNPY2 canopy FGF signaling regulator 2 4 108 O14773 1200 TPP1 tripeptidyl peptidase I 2 109 Q9UNW1 9562 MINPP1 multiple inositol-polyphosphate phosphatase 1 4 110 P02749 350 APOH apolipoprotein H 3 111 P13667 9601 PDIA4 protein disulfide isomerase family A member 4 8 112 Q03167 7049 TGFBR3 transforming growth factor beta receptor III 2 113 P01031 727 C5 complement component 5 11 114 P14543 4811 NID1 nidogen 1 4 115 Q9BWS9 66005 CHID1 chitinase domain containing 1 3 116 Q99574 5274 SERPINI1 serpin peptidase inhibitor, clade I (neuroserpin), member 1 2 117 Q14978 9221 NOLC1 nucleolar and coiled-body phosphoprotein 1 2 118 Q06033 3699 ITIH3 inter-alpha-trypsin inhibitor heavy chain 3 5 119 Q96HE7 30001 ERO1A endoplasmic reticulum oxidoreductase alpha 6 120 Q14554 10954 PDIA5 protein disulfide isomerase family A member 5 2 121 P23141 1066 CES1 carboxylesterase 1 2 122 O94985 22883 CLSTN1 calsyntenin 1 2 123 P07996 7057 THBS1 thrombospondin 1 3 124 P02545 4000 LMNA lamin A/C 2 125 P10253 2548 GAA glucosidase, alpha; acid 2 126 P15144 290 ANPEP alanyl aminopeptidase, membrane 5 127 P35613 682 BSG basigin (Ok blood group) 2 128 O00748 8824 CES2 carboxylesterase 2 4 129 Q02809 5351 PLOD1 procollagen-lysine, 2-oxoglutarate 5-dioxygenase 1 2 130 P61160 10097 ACTR2 ARP2 actin-related protein 2 homolog (yeast) 2 131 P55145 7873 MANF mesencephalic astrocyte derived neurotrophic factor 4 132 P11021 3309 HSPA5 heat shock protein family A (Hsp70) member 5 2 133 P12956 2547 XRCC6 X-ray repair complementing defective repair in Chinese hamster cells 6 2 134 P55157 4547 MTTP microsomal triglyceride transfer protein 2 135 Q9H488 23509 POFUT1 protein O-fucosyltransferase 1 4 136 P00738 3240 HP haptoglobin 2 137 P62158 801 CALM1 calmodulin 1 (phosphorylase kinase, delta) 2 138 P05156 3426 CFI complement factor I 6 139 P16870 1363 CPE carboxypeptidase E 2 140 P62993 2885 GRB2 growth factor receptor bound protein 2 3 141 P22307 6342 SCP2 sterol carrier protein 2 2 142 P15586 2799 GNS glucosamine (N-acetyl)-6-sulfatase 2 143 P23284 5479 PPIB peptidylprolyl isomerase B 2 144 P00742 2159 F10 coagulation factor X 2 145 P06576 506 ATP5B ATP synthase, H+ transporting, mitochondrial F1 complex, beta polypeptide 7

<2-4> Cell function analysis of secreted proteins showing expression change according to oleate treatment

Among the identified proteins showing a change in expression level by oleate treatment in <2-3>, functional analysis was performed using DAVID software to identify GOBPs and KEGG pathways representing 145 proteins with increased secretion. Was performed.

As a result, as shown in FIG. 6, the proteins increased during oleate treatment were significantly involved in cell functions as shown in Table 2 below.

Cell function mainly related to proteins with increased secretion by oleate treatment number Description Count P value One extracellular matrix organization 21 1.82E-10 2 protein folding 17 2.89E-10 3 response to wounding 33 6.00E-10 4 proteolysis 40 2.17E-09 5 lipid transport 15 3.44E-07 6 cell migration 26 2.46E-05 7 endocytosis 18 3.61E-05 8 inflammatory response 17 1.09E-04 9 lipid metabolic process 23 0.001905 10 cell adhesion 24 0.003426 11 Lysosome 12 5.01E-07 12 Protein processing in endoplasmic reticulum 12 1.20E-05 13 Complement and coagulation cascades 8 3.31E-05

<2-5> Identification of new protein showing up-regulation by oleate treatment in liver cancer cell line

Among the total of 145 proteins in which the expression level is increased upon oleate treatment in the liver cancer cell line, there is no report on the relationship with the conventional liver cancer and the expression and secretion in liver cancer cells are increased compared to the group not treated with oleate treatment. 18 proteins were identified, and the protein list is shown in Table 3 below.

List of new proteins that increase in liver cancer cells during oleate treatment number UniProtKB
accession Entrez ID Symbol Description log2-fold
change Sequence number
(amino acid) One Q96DT5 8701 DNAH11 dynein, axonemal, heavy chain 11 2.865 One 2 Q9BPW8 8508 NIPSNAP1 nipsnap homolog 1 1.719 2 3 O15067 5198 PFAS phosphoribosylformylglycinamidine synthase 1.696 3 4 Q6IN85 55671 PPP4R3A protein phosphatase 4 regulatory subunit 3A 1.527 4 5 Q8NI22 90411 MCFD2 multiple coagulation factor deficiency 2 1.375 5 6 O43847 4898 NRDC nardilysin convertase 1.354 6 7 Q86V81 10189 ALYREF Aly/REF export factor 1.291 7 8 P51659 3295 HSD17B4 hydroxysteroid (17-beta) dehydrogenase 4 1.265 8 9 Q14651 5357 PLS1 plastin 1 1.212 9 10 P20142 5225 PGC progastricsin (pepsinogen C) 1.165 10 11 Q9H993 79624 ARMT1 acidic residue methyltransferase 1 1.099 11 12 Q9UHG2 27344 PCSK1N proprotein convertase subtilisin/kexin type 1 inhibitor 1.070 12 13 Q9NYU2 56886 UGGT1 UDP-glucose glycoprotein glucosyltransferase 1 1.034 13 14 O95433 10598 AHSA1 AHA1, activator of heat shock 90kDa protein ATPase homolog 1 (yeast) 1.031 14 15 P02795 4502 MT2A metallothionein 2A 1.029 15 16 Q14696 23184 MESDC2 mesoderm development candidate 2 1.016 16 17 P06865 3073 HEXA hexosaminidase subunit alpha 1.014 17 18 P19823 3698 ITIH2 inter-alpha-trypsin inhibitor heavy chain 2 1.002 18

Through the above results, the present inventors found that the 18 proteins in Table 3 can be used as a new biomarker for liver cancer diagnosis, and can also be used as a biomarker to confirm whether cancer cells proliferate by oleate. Was found.

So far, the present invention has been focused on the preferred embodiments. Those skilled in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered in terms of explanation, not limitation. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the equivalent range should be interpreted as being included in the present invention.

<110> Korea Brain Research Institute <120> Novel hepatocellular carcinomar diagnostic marker and use thereof <130> NPDC-71084 <160> 18 <170> KoPatentIn 3.0 <210> 1 <211> 4516 <212> PRT <213> Artificial Sequence <220> <223> DNAH11 amino acid sequence <400> 1 Met Ala Ala Gln Val Ala Ala Arg Glu Ala Arg Asp Phe Arg Glu Ala 1 5 10 15 Pro Thr Leu Arg Leu Thr Ser Gly Ala Gly Leu Glu Ala Val Gly Ala 20 25 30 Val Glu Leu Glu Glu Glu Glu Glu Asn Glu Glu Glu Ala Ala Ala Arg 35 40 45 Arg Ala Arg Ser Phe Ala Gln Asp Ala Arg Val Arg Phe Leu Gly Gly 50 55 60 Arg Leu Ala Met Met Leu Gly Phe Thr Glu Glu Lys Trp Ser Gln Tyr 65 70 75 80 Leu Glu Ser Glu Asp Asn Arg Gln Val Leu Gly Glu Phe Leu Glu Ser 85 90 95 Thr Ser Pro Ala Cys Leu Val Phe Ser Phe Ala Ala Ser Gly Arg Leu 100 105 110 Ala Ala Ser Gln Glu Ile Pro Arg Asp Ala Asn His Lys Leu Val Phe 115 120 125 Ile Ser Lys Lys Ile Thr Glu Ser Ile Gly Val Asn Asp Phe Ser Gln 130 135 140 Val Val Leu Phe Gly Glu Leu Pro Ala Leu Ser Leu Gly His Val Ser 145 150 155 160 Ala Phe Leu Asp Glu Ile Leu Val Pro Val Leu Ser Asn Lys Asn Asn 165 170 175 His Lys Ser Trp Ser Cys Phe Thr Ser Gln Asp Met Glu Tyr His Ile 180 185 190 Glu Val Met Lys Lys Lys Met Tyr Ile Phe Arg Gly Lys Met Ser Arg 195 200 205 Arg Thr Leu Leu Pro Ile Pro Thr Val Ala Gly Lys Met Asp Leu Asp 210 215 220 Gln Asn Cys Ser Glu Asn Lys Pro Pro Ser Asn Glu Arg Ile Ile Leu 225 230 235 240 His Ala Ile Glu Ser Val Val Ile Glu Trp Ser His Gln Ile Gln Glu 245 250 255 Ile Ile Glu Arg Asp Ser Val Gln Arg Leu Leu Asn Gly Leu His Leu 260 265 270 Ser Pro Gln Ala Glu Leu Asp Phe Trp Met Met Arg Arg Glu Asn Leu 275 280 285 Ser Cys Ile Tyr Asp Gln Leu Gln Ala Pro Val Val Leu Lys Met Val 290 295 300 Lys Ile Leu Thr Thr Lys Gln Ser Ser Tyr Phe Pro Thr Leu Lys Asp 305 310 315 320 Ile Phe Leu Ala Val Glu Asn Ala Leu Leu Glu Ala Gln Asp Val Glu 325 330 335 Leu Tyr Leu Arg Pro Leu Arg Arg His Ile Gln Cys Leu Gln Glu Thr 340 345 350 Glu Phe Pro Gln Thr Arg Ile Leu Ile Ala Pro Leu Phe His Thr Ile 355 360 365 Cys Leu Ile Trp Ser His Ser Lys Phe Tyr Asn Thr Pro Ala Arg Val 370 375 380 Ile Val Leu Leu Gln Glu Phe Cys Asn Leu Phe Ile Asn Gln Ala Thr 385 390 395 400 Ala Tyr Leu Ser Pro Glu Asp Leu Leu Arg Gly Glu Ile Glu Glu Ser 405 410 415 Leu Glu Lys Val Gln Val Ala Val Asn Ile Leu Lys Thr Phe Lys Asn 420 425 430 Ser Phe Phe Asn Tyr Arg Lys Lys Leu Ala Ser Tyr Phe Met Gly Arg 435 440 445 Lys Leu Arg Pro Trp Asp Phe Gln Ser His Leu Val Phe Cys Arg Phe 450 455 460 Asp Lys Phe Leu Asp Arg Leu Ile Lys Ile Glu Asp Ile Phe Ala Thr 465 470 475 480 Thr Leu Glu Phe Glu Lys Leu Glu Arg Leu Glu Phe Gly Gly Thr Lys 485 490 495 Gly Ala Ile Leu Asn Gly Gln Val His Glu Met Ser Glu Glu Leu Met 500 505 510 Glu Leu Cys Lys Leu Phe Lys Gln Ser Thr Tyr Asp Pro Ser Asp Cys 515 520 525 Thr Asn Met Glu Phe Glu Ser Asp Tyr Val Ala Phe Lys Ser Lys Thr 530 535 540 Leu Glu Phe Asp Arg Arg Leu Gly Thr Ile Ile Cys Glu Ala Phe Phe 545 550 555 560 Asn Cys Asn Gly Leu Glu Ala Ala Phe Lys Leu Leu Thr Ile Phe Gly 565 570 575 Asn Phe Leu Glu Lys Pro Val Val Met Glu Ile Phe Ser Leu His Tyr 580 585 590 Ser Thr Leu Val His Met Phe Asn Thr Glu Leu Asp Val Cys Lys Gln 595 600 605 Leu Tyr Asn Glu His Met Lys Gln Ile Glu Cys Gly His Val Val Leu 610 615 620 Asn Lys Asn Met Pro Phe Thr Ser Gly Asn Met Lys Trp Ala Gln Gln 625 630 635 640 Val Leu Gln Arg Leu Gln Met Phe Trp Ser Asn Phe Ala Ser Leu Arg 645 650 655 Tyr Leu Phe Leu Gly Asn Pro Asp His Ala Leu Val Tyr Gln Lys Tyr 660 665 670 Val Glu Met Thr Thr Leu Leu Asp Gln Phe Glu Ser Arg Ile Tyr Asn 675 680 685 Glu Trp Lys Ser Asn Val Asp Glu Ile Cys Glu Phe Asn Leu Asn Gln 690 695 700 Pro Leu Val Lys Phe Ser Ala Ile Asn Gly Leu Leu Cys Val Asn Phe 705 710 715 720 Asp Pro Lys Leu Val Ala Val Leu Arg Glu Val Lys Tyr Leu Leu Met 725 730 735 Leu Lys Lys Gln Asp Ile Pro Asp Ser Ala Leu Ala Ile Phe Lys Lys 740 745 750 Arg Asn Thr Ile Leu Lys Tyr Ile Gly Asn Leu Asp Leu Leu Val Gln 755 760 765 Gly Tyr Asn Lys Leu Lys Gln Thr Leu Leu Glu Val Glu Tyr Pro Leu 770 775 780 Ile Glu Asp Glu Leu Arg Ala Ile Asp Glu Gln Leu Thr Ala Ala Thr 785 790 795 800 Thr Trp Leu Thr Trp Gln Asp Asp Cys Trp Gly Tyr Ile Glu Arg Val 805 810 815 Arg Ala Ala Thr Ser Glu Leu Glu His Arg Val Glu Arg Thr Gln Lys 820 825 830 Asn Val Lys Val Ile Gln Gln Thr Met Arg Gly Trp Ala Arg Cys Val 835 840 845 Leu Pro Pro Arg Arg Glu His Arg Arg Glu Ala Ala Phe Thr Leu Glu 850 855 860 Asp Lys Gly Asp Leu Phe Thr Lys Lys Tyr Lys Leu Ile Gln Gly Asp 865 870 875 880 Gly Cys Lys Ile His Asn Leu Val Glu Glu Asn Arg Lys Leu Phe Lys 885 890 895 Ala Asn Pro Ser Leu Asp Thr Trp Lys Ile Tyr Val Glu Phe Ile Asp 900 905 910 Asp Ile Val Val Glu Gly Phe Phe Gln Ala Ile Met His Asp Leu Asp 915 920 925 Phe Phe Leu Lys Asn Thr Glu Lys Gln Leu Lys Pro Ala Pro Phe Phe 930 935 940 Gln Ala Gln Met Ile Leu Leu Pro Pro Glu Ile Val Phe Lys Pro Ser 945 950 955 960 Leu Asp Arg Glu Ala Gly Asp Gly Phe Tyr Asp Leu Val Glu Glu Met 965 970 975 Leu Cys Asn Ser Phe Arg Met Ser Ala Gln Met Asn Arg Ile Ala Thr 980 985 990 His Leu Glu Ile Lys Asn Tyr Gln Asn Asp Met Asp Asn Met Leu Gly 995 1000 1005 Leu Ala Glu Val Arg Gln Glu Ile Met Asn Arg Val Val Asn Val Ile 1010 1015 1020 Asn Lys Val Leu Asp Phe Arg Asn Thr Leu Glu Thr His Thr Tyr Leu 1025 1030 1035 1040 Trp Val Asp Asp Arg Ala Glu Phe Met Lys His Phe Leu Leu Tyr Gly 1045 1050 1055 His Ala Val Ser Ser Asp Glu Met Asp Ala His Ala Asn Glu Glu Ile 1060 1065 1070 Pro Glu Gln Pro Pro Thr Leu Glu Gln Phe Lys Glu Gln Ile Asp Ile 1075 1080 1085 Tyr Glu Ala Leu Tyr Val Gln Met Ser Lys Phe Glu Asp Phe Arg Val 1090 1095 1100 Phe Asp Ser Trp Phe Lys Val Asp Met Lys Pro Phe Lys Val Ser Leu 1105 1110 1115 1120 Leu Thr Ile Ile Lys Lys Trp Ser Trp Met Phe Gln Glu His Leu Leu 1125 1130 1135 Arg Phe Val Ile Asp Ser Leu Asn Glu Leu Gln Glu Phe Ile Lys Glu 1140 1145 1150 Thr Asp Ser Gly Leu Gln Arg Glu Leu Asn Glu Gly Asp His Asp Gly 1155 1160 1165 Leu Val Asp Ile Met Val His Leu Leu Ala Val Arg Ser Arg Gln Arg 1170 1175 1180 Ala Thr Asp Glu Leu Phe Glu Pro Leu Lys Glu Thr Ile Thr Leu Leu 1185 1190 1195 1200 Glu Ser Tyr Gly Gln Lys Met Pro Glu Gln Val Tyr Ile Gln Leu Glu 1205 1210 1215 Glu Leu Pro Glu Arg Trp Glu Thr Thr Lys Lys Ile Ala Ala Thr Val 1220 1225 1230 Arg His Glu Val Ser Pro Leu His Asn Ala Glu Val Thr Leu Ile Arg 1235 1240 1245 Lys Lys Cys Ile Leu Phe Asp Ala Lys Gln Ala Glu Phe Arg Glu Arg 1250 1255 1260 Phe Arg His Tyr Ala Pro Leu Gly Phe Asn Ala Glu Asn Pro Tyr Thr 1265 1270 1275 1280 Ala Leu Asp Lys Ala Asn Glu Glu Leu Glu Ala Leu Glu Glu Glu Met 1285 1290 1295 Leu Gln Met Gln Glu Ser Thr Arg Leu Phe Glu Val Ala Leu Pro Glu 1300 1305 1310 Tyr Lys Gln Met Lys Gln Cys Arg Lys Glu Ile Lys Leu Leu Lys Gly 1315 1320 1325 Leu Trp Asp Val Ile Ile Tyr Val Arg Arg Ser Ile Asp Asn Trp Thr 1330 1335 1340 Lys Thr Gln Trp Arg Gln Ile His Val Glu Gln Met Asp Val Glu Leu 1345 1350 1355 1360 Arg Arg Phe Ala Lys Glu Ile Trp Ser Leu Asn Lys Glu Val Arg Val 1365 1370 1375 Trp Asp Ala Tyr Thr Gly Leu Glu Gly Thr Val Lys Asp Met Thr Ala 1380 1385 1390 Ser Leu Arg Ala Ile Thr Glu Leu Gln Ser Pro Ala Leu Arg Asp Arg 1395 1400 1405 His Trp His Gln Leu Met Lys Ala Ile Gly Val Lys Phe Leu Ile Asn 1410 1415 1420 Glu Ala Thr Thr Leu Ala Asp Leu Leu Ala Leu Arg Leu His Arg Val 1425 1430 1435 1440 Glu Asp Asp Val Arg Arg Ile Val Asp Lys Ala Val Lys Glu Leu Gly 1445 1450 1455 Thr Glu Lys Val Ile Thr Glu Ile Ser Gln Thr Trp Ala Thr Met Lys 1460 1465 1470 Phe Ser Tyr Glu Val His Tyr Arg Thr Gly Ile Pro Leu Leu Lys Ser 1475 1480 1485 Asp Glu Gln Leu Phe Glu Thr Leu Glu His Asn Gln Val Gln Leu Gln 1490 1495 1500 Thr Leu Leu Gln Ser Lys Tyr Val Glu Tyr Phe Ile Glu Gln Val Leu 1505 1510 1515 1520 Ser Trp Gln Asn Lys Leu Asn Ile Ala Asp Leu Val Ile Phe Thr Trp 1525 1530 1535 Met Glu Val Gln Arg Thr Trp Ser His Leu Glu Ser Ile Phe Val Cys 1540 1545 1550 Ser Glu Asp Ile Arg Ile Gln Leu Val Lys Asp Ala Arg Arg Phe Asp 1555 1560 1565 Gly Val Asp Ala Glu Phe Lys Glu Leu Met Phe Lys Thr Ala Lys Val 1570 1575 1580 Glu Asn Val Leu Glu Ala Thr Cys Arg Pro Asn Leu Tyr Glu Lys Leu 1585 1590 1595 1600 Lys Asp Leu Gln Ser Arg Leu Ser Leu Cys Glu Lys Ala Leu Ala Glu 1605 1610 1615 Tyr Leu Glu Thr Lys Arg Ile Ala Phe Pro Arg Phe Tyr Phe Val Ser 1620 1625 1630 Ser Ala Asp Leu Leu Asp Ile Leu Ser Lys Gly Ala Gln Pro Lys Gln 1635 1640 1645 Val Thr Cys His Leu Ala Lys Leu Phe Asp Ser Ile Ala Asp Leu Gln 1650 1655 1660 Phe Glu Asp Asn Gln Asp Val Ser Ala His Arg Ala Val Gly Met Tyr 1665 1670 1675 1680 Ser Lys Glu Lys Glu Tyr Val Pro Phe Gln Ala Glu Cys Glu Cys Val 1685 1690 1695 Gly His Val Glu Thr Trp Leu Leu Gln Leu Glu Gln Thr Met Gln Glu 1700 1705 1710 Thr Val Arg His Ser Ile Thr Glu Ala Ile Val Ala Tyr Glu Glu Lys 1715 1720 1725 Pro Arg Glu Leu Trp Ile Phe Asp Phe Pro Ala Gln Val Ala Leu Thr 1730 1735 1740 Ser Ser Gln Ile Trp Trp Thr Thr Asp Val Gly Ile Ala Phe Ser Arg 1745 1750 1755 1760 Leu Glu Glu Gly Tyr Glu Thr Ala Leu Lys Asp Phe His Lys Lys Gln 1765 1770 1775 Ile Ser Gln Leu Asn Thr Leu Ile Thr Leu Leu Leu Gly Glu Leu Pro 1780 1785 1790 Pro Gly Asp Arg Gln Lys Ile Met Thr Ile Cys Thr Ile Asp Val His 1795 1800 1805 Ala Arg Asp Val Val Ala Lys Leu Ile Ser Gln Lys Val Val Ser Pro 1810 1815 1820 Gln Ala Phe Thr Trp Leu Ser Gln Leu Arg His Arg Trp Glu Asp Thr 1825 1830 1835 1840 Gln Lys His Cys Phe Val Asn Ile Cys Asp Ala Gln Phe Gln Tyr Phe 1845 1850 1855 Tyr Glu Tyr Leu Gly Asn Ser Pro Arg Leu Val Ile Thr Pro Leu Thr 1860 1865 1870 Asp Arg Cys Tyr Ile Thr Leu Thr Gln Ser Leu His Leu Thr Met Ser 1875 1880 1885 Gly Ala Pro Ala Gly Pro Ala Gly Thr Gly Lys Thr Glu Thr Thr Lys 1890 1895 1900 Asp Leu Gly Arg Ala Leu Gly Met Met Val Tyr Val Phe Asn Cys Ser 1905 1910 1915 1920 Glu Gln Met Asp Tyr Lys Ser Ile Gly Asn Ile Tyr Lys Gly Leu Val 1925 1930 1935 Gln Thr Gly Ala Trp Gly Cys Phe Asp Glu Phe Asn Arg Ile Ser Val 1940 1945 1950 Glu Val Leu Ser Val Val Ala Val Gln Val Lys Met Ile His Asp Ala 1955 1960 1965 Ile Arg Asn Arg Lys Lys Arg Phe Val Phe Leu Gly Glu Ala Ile Thr 1970 1975 1980 Leu Lys Pro Ser Val Gly Ile Phe Ile Thr Met Asn Pro Gly Tyr Ala 1985 1990 1995 2000 Gly Arg Thr Glu Leu Pro Glu Asn Leu Lys Ala Leu Phe Arg Pro Cys 2005 2010 2015 Ala Met Val Ala Pro Asp Ile Glu Leu Ile Cys Glu Ile Leu Leu Val 2020 2025 2030 Ala Glu Gly Phe Val Asp Ala Arg Ala Leu Ala Arg Lys Phe Ile Thr 2035 2040 2045 Leu Tyr Thr Leu Cys Lys Glu Leu Leu Ser Lys Gln Asp His Tyr Asp 2050 2055 2060 Trp Gly Leu Arg Ala Ile Lys Ser Val Leu Val Val Ala Gly Ser Leu 2065 2070 2075 2080 Lys Arg Gly Asp Lys Asn Arg Pro Glu Asp Gln Val Leu Met Arg Ala 2085 2090 2095 Leu Arg Asp Phe Asn Met Pro Lys Ile Val Thr Asp Asp Ile Pro Val 2100 2105 2110 Phe Leu Gly Leu Val Gly Asp Leu Phe Pro Ala Leu Asp Val Pro Arg 2115 2120 2125 Arg Arg Lys Leu His Phe Glu Gln Met Val Arg Gln Ser Thr Leu Glu 2130 2135 2140 Leu Arg Leu Gln Pro Glu Glu Ser Phe Ile Leu Lys Val Val Gln Leu 2145 2150 2155 2160 Glu Glu Leu Leu Ala Val Arg His Ser Val Phe Val Val Gly Asn Ala 2165 2170 2175 Gly Thr Gly Lys Ser Lys Ile Leu Arg Thr Leu Asn Arg Thr Tyr Val 2180 2185 2190 Asn Met Lys Gln Lys Pro Val Trp Asn Asp Leu Asn Pro Lys Ala Val 2195 2200 2205 Thr Thr Asp Glu Leu Phe Gly Phe Ile His His Ala Thr Arg Glu Trp 2210 2215 2220 Lys Asp Gly Lys Ile Val Tyr Ser Tyr Phe Ile Gly Leu Phe Ser Ser 2225 2230 2235 2240 Ile Leu Arg Glu Gln Ala Asn Leu Lys His Asp Gly Pro Lys Trp Ile 2245 2250 2255 Val Leu Asp Gly Asp Ile Asp Pro Met Trp Ile Glu Ser Leu Asn Thr 2260 2265 2270 Val Met Asp Asp Asn Lys Val Leu Thr Leu Ala Ser Asn Glu Arg Ile 2275 2280 2285 Ala Leu Thr Pro Phe Met Arg Leu Leu Phe Glu Ile His His Leu Arg 2290 2295 2300 Ser Ala Thr Pro Ala Thr Val Ser Arg Ala Gly Ile Leu Tyr Val Asn 2305 2310 2315 2320 Pro Gln Asp Leu Gly Trp Asn Pro Tyr Val Ala Ser Trp Ile Asp Arg 2325 2330 2335 Arg Arg His Gln Ser Glu Lys Ala Asn Leu Thr Ile Leu Phe Asp Lys 2340 2345 2350 Tyr Val Pro Ala Cys Leu Asp Lys Leu Arg Thr Ser Phe Lys Thr Ile 2355 2360 2365 Thr Ser Ile Pro Glu Ser Ser Leu Val Gln Thr Leu Cys Val Leu Leu 2370 2375 2380 Glu Cys Leu Leu Thr Pro Glu Asn Val Pro Ser Asp Ser Pro Lys Glu 2385 2390 2395 2400 Val Tyr Glu Val Tyr Phe Val Phe Ala Cys Ile Trp Ala Phe Gly Gly 2405 2410 2415 Thr Leu Leu Gln Asp Gln Ile Ser Asp Tyr Gln Ala Asp Phe Ser Arg 2420 2425 2430 Trp Trp Gln Lys Glu Met Lys Ala Val Lys Phe Pro Ser Gln Gly Thr 2435 2440 2445 Ile Phe Asp Tyr Tyr Val Asp His Lys Thr Lys Lys Leu Leu Pro Trp 2450 2455 2460 Ala Asp Lys Ile Ala Gln Phe Thr Met Asp Pro Asp Val Pro Leu Gln 2465 2470 2475 2480 Thr Val Leu Val His Thr Thr Glu Thr Ala Arg Leu Arg Tyr Phe Met 2485 2490 2495 Glu Leu Leu Leu Glu Lys Gly Lys Pro Leu Met Leu Val Gly Asn Ala 2500 2505 2510 Gly Val Gly Lys Thr Val Phe Val Gly Asp Thr Leu Ala Ser Leu Ser 2515 2520 2525 Glu Asp Tyr Ile Val Ser Arg Val Pro Phe Asn Tyr Tyr Thr Thr Ser 2530 2535 2540 Thr Ala Leu Gln Lys Ile Leu Glu Lys Pro Leu Glu Lys Lys Ala Gly 2545 2550 2555 2560 His Asn Tyr Gly Pro Gly Gly Asn Lys Lys Leu Ile Tyr Phe Ile Asp 2565 2570 2575 Asp Met Asn Met Pro Glu Val Asp Leu Tyr Gly Thr Val Gln Pro His 2580 2585 2590 Thr Leu Ile Arg Gln His Ile Asp Tyr Gly His Trp Tyr Asp Arg Gln 2595 2600 2605 Lys Val Met Leu Lys Glu Ile His Asn Cys Gln Tyr Val Ala Cys Met 2610 2615 2620 Asn Pro Met Val Gly Ser Phe Thr Ile Asn Pro Arg Leu Gln Arg His 2625 2630 2635 2640 Phe Thr Val Phe Ala Phe Asn Phe Pro Ser Leu Asp Ala Leu Asn Thr 2645 2650 2655 Ile Tyr Gly Gln Ile Phe Ser Phe His Phe Gln Gln Gln Ala Phe Ala 2660 2665 2670 Pro Ser Ile Leu Arg Ser Gly Pro Thr Leu Ile Gln Ala Thr Ile Ala 2675 2680 2685 Phe His Gln Thr Met Met Cys Asn Phe Leu Pro Thr Ala Ile Lys Phe 2690 2695 2700 His Tyr Ile Phe Asn Leu Arg Asp Leu Ser Asn Val Phe Gln Gly Ile 2705 2710 2715 2720 Leu Phe Ala Ser Pro Glu Cys Leu Lys Gly Pro Leu Asp Leu Ile His 2725 2730 2735 Leu Trp Leu His Glu Ser Ala Arg Val Tyr Gly Asp Lys Leu Ile Asp 2740 2745 2750 Lys Lys Asp Cys Asp Leu Phe Gln Arg Arg Met Leu Glu Thr Ala Tyr 2755 2760 2765 Lys Tyr Phe Glu Gly Ile Asp Ser His Met Leu Leu Gln Gln Pro Leu 2770 2775 2780 Ile Tyr Cys His Phe Ala Asp Arg Gly Lys Asp Pro His Tyr Met Pro 2785 2790 2795 2800 Val Lys Asp Trp Glu Val Leu Lys Thr Ile Leu Thr Glu Thr Leu Asp 2805 2810 2815 Asn Tyr Asn Glu Leu Asn Ala Ala Met His Leu Val Leu Phe Glu Asp 2820 2825 2830 Ala Met Gln His Val Cys Arg Ile Ser Arg Ile Leu Arg Thr Pro Gln 2835 2840 2845 Gly Cys Ala Leu Leu Val Gly Val Gly Gly Ser Gly Lys Gln Ser Leu 2850 2855 2860 Ser Arg Leu Ala Ala Tyr Leu Arg Gly Leu Glu Val Phe Gln Ile Thr 2865 2870 2875 2880 Leu Thr Glu Gly Tyr Gly Ile Gln Glu Leu Arg Val Asp Leu Ala Asn 2885 2890 2895 Leu Tyr Ile Arg Thr Gly Ala Lys Asn Met Pro Thr Val Phe Leu Leu 2900 2905 2910 Thr Asp Ala Gln Val Leu Asp Glu Ser Phe Leu Val Leu Ile Asn Asp 2915 2920 2925 Leu Leu Ala Ser Gly Glu Ile Pro Asp Leu Phe Ser Asp Glu Asp Val 2930 2935 2940 Asp Lys Ile Ile Ser Gly Ile His Asn Glu Val His Ala Leu Gly Met 2945 2950 2955 2960 Val Asp Ser Arg Glu Asn Cys Trp Lys Phe Phe Met Ala Arg Val Arg 2965 2970 2975 Leu Gln Leu Lys Ile Ile Leu Cys Phe Ser Pro Val Gly Arg Thr Leu 2980 2985 2990 Arg Val Arg Ala Arg Lys Phe Pro Ala Ile Val Asn Cys Thr Ala Ile 2995 3000 3005 Asp Trp Phe His Ala Trp Pro Gln Glu Ala Leu Val Ser Val Ser Arg 3010 3015 3020 Arg Phe Ile Glu Glu Thr Lys Gly Ile Glu Pro Val His Lys Asp Ser 3025 3030 3035 3040 Ile Ser Leu Phe Met Ala His Val His Thr Thr Val Asn Glu Met Ser 3045 3050 3055 Thr Arg Tyr Tyr Gln Asn Glu Arg Arg His Asn Tyr Thr Thr Pro Lys 3060 3065 3070 Ser Phe Leu Glu Gln Ile Ser Leu Phe Lys Asn Leu Leu Lys Lys Lys 3075 3080 3085 Gln Asn Glu Val Ser Glu Lys Lys Glu Arg Leu Val Asn Gly Ile Gln 3090 3095 3100 Lys Leu Lys Thr Thr Ala Ser Gln Val Gly Asp Leu Lys Ala Arg Leu 3105 3110 3115 3120 Ala Ser Gln Glu Ala Glu Leu Gln Leu Arg Asn His Asp Ala Glu Ala 3125 3130 3135 Leu Ile Thr Lys Ile Gly Leu Gln Thr Glu Lys Val Ser Arg Glu Lys 3140 3145 3150 Thr Ile Ala Asp Ala Glu Glu Arg Lys Val Thr Ala Ile Gln Thr Glu 3155 3160 3165 Val Phe Gln Lys Gln Arg Glu Cys Glu Ala Asp Leu Leu Lys Ala Glu 3170 3175 3180 Pro Ala Leu Val Ala Ala Thr Ala Ala Leu Asn Thr Leu Asn Arg Val 3185 3190 3195 3200 Asn Leu Ser Glu Leu Lys Ala Phe Pro Asn Pro Pro Ile Ala Val Thr 3205 3210 3215 Asn Val Thr Ala Ala Val Met Val Leu Leu Ala Pro Arg Gly Arg Val 3220 3225 3230 Pro Lys Asp Arg Ser Trp Lys Ala Ala Lys Val Phe Met Gly Lys Val 3235 3240 3245 Asp Asp Phe Leu Gln Ala Leu Ile Asn Tyr Asp Lys Glu His Ile Pro 3250 3255 3260 Glu Asn Cys Leu Lys Val Val Asn Glu His Tyr Leu Lys Asp Pro Glu 3265 3270 3275 3280 Phe Asn Pro Asn Leu Ile Arg Thr Lys Ser Phe Ala Ala Ala Gly Leu 3285 3290 3295 Cys Ala Trp Val Ile Asn Ile Ile Lys Phe Tyr Glu Val Tyr Cys Asp 3300 3305 3310 Val Glu Pro Lys Arg Gln Ala Leu Ala Gln Ala Asn Leu Glu Leu Ala 3315 3320 3325 Ala Ala Thr Glu Lys Leu Glu Ala Ile Arg Lys Lys Leu Val Asp Leu 3330 3335 3340 Asp Arg Asn Leu Ser Arg Leu Thr Ala Ser Phe Glu Lys Ala Thr Ala 3345 3350 3355 3360 Glu Lys Val Arg Cys Gln Glu Glu Val Asn Gln Thr Asn Lys Thr Ile 3365 3370 3375 Lys Leu Ala Asn Arg Leu Val Lys Glu Leu Glu Ala Lys Lys Ile Arg 3380 3385 3390 Trp Gly Gln Ser Ile Lys Ser Phe Glu Ala Gln Glu Lys Thr Leu Cys 3395 3400 3405 Gly Asp Val Leu Leu Thr Ala Ala Phe Val Ser Tyr Val Gly Pro Phe 3410 3415 3420 Thr Arg Gln Tyr Arg Gln Glu Leu Val His Cys Lys Trp Val Pro Phe 3425 3430 3435 3440 Leu Gln Gln Lys Val Ser Ile Pro Leu Thr Glu Gly Leu Asp Leu Ile 3445 3450 3455 Ser Met Leu Thr Asp Asp Ala Thr Ile Ala Ala Trp Asn Asn Glu Gly 3460 3465 3470 Leu Pro Ser Asp Arg Met Ser Thr Glu Asn Ala Ala Ile Leu Thr His 3475 3480 3485 Cys Glu Arg Trp Pro Leu Val Ile Asp Pro Gln Gln Gln Gly Ile Lys 3490 3495 3500 Trp Ile Lys Asn Lys Tyr Gly Met Asp Leu Lys Val Thr His Leu Gly 3505 3510 3515 3520 Gln Lys Gly Phe Leu Asn Ala Ile Glu Thr Ala Leu Ala Phe Gly Asp 3525 3530 3535 Val Ile Leu Ile Glu Asn Leu Glu Glu Thr Ile Asp Pro Val Leu Asp 3540 3545 3550 Pro Leu Leu Gly Arg Asn Thr Ile Lys Lys Gly Lys Tyr Ile Arg Ile 3555 3560 3565 Gly Asp Lys Glu Cys Glu Phe Asn Lys Asn Phe Arg Leu Ile Leu His 3570 3575 3580 Thr Lys Leu Ala Asn Pro His Tyr Lys Pro Glu Leu Gln Ala Gln Thr 3585 3590 3595 3600 Thr Leu Leu Asn Phe Thr Val Thr Glu Asp Gly Leu Glu Ala Gln Leu 3605 3610 3615 Leu Ala Glu Val Val Ser Ile Glu Arg Pro Asp Leu Glu Lys Leu Lys 3620 3625 3630 Leu Val Leu Thr Lys His Gln Asn Asp Phe Lys Ile Glu Leu Lys Tyr 3635 3640 3645 Leu Glu Asp Asp Leu Leu Leu Arg Leu Ser Ala Ala Glu Gly Ser Phe 3650 3655 3660 Leu Asp Asp Thr Lys Leu Val Glu Arg Leu Glu Ala Thr Lys Thr Thr 3665 3670 3675 3680 Val Ala Glu Ile Glu His Lys Val Ile Glu Ala Lys Glu Asn Glu Arg 3685 3690 3695 Lys Ile Asn Glu Ala Arg Glu Cys Tyr Arg Pro Val Ala Ala Arg Ala 3700 3705 3710 Ser Leu Leu Tyr Phe Val Ile Asn Asp Leu Gln Lys Ile Asn Pro Leu 3715 3720 3725 Tyr Gln Phe Ser Leu Lys Ala Phe Asn Val Leu Phe His Arg Ala Ile 3730 3735 3740 Glu Gln Ala Asp Lys Val Glu Asp Met Gln Gly Arg Ile Ser Ile Leu 3745 3750 3755 3760 Met Glu Ser Ile Thr His Ala Val Phe Leu Tyr Thr Ser Gln Ala Leu 3765 3770 3775 Phe Glu Lys Asp Lys Leu Thr Phe Leu Ser Gln Met Ala Phe Gln Ile 3780 3785 3790 Leu Leu Arg Lys Lys Glu Ile Asp Pro Leu Glu Leu Asp Phe Leu Leu 3795 3800 3805 Arg Phe Thr Val Glu His Thr His Leu Ser Pro Val Asp Phe Leu Thr 3810 3815 3820 Ser Gln Ser Trp Ser Ala Ile Lys Ala Ile Ala Val Met Glu Glu Phe 3825 3830 3835 3840 Arg Gly Ile Asp Arg Asp Val Glu Gly Ser Ala Lys Gln Trp Arg Lys 3845 3850 3855 Trp Val Glu Ser Glu Cys Pro Glu Lys Glu Lys Leu Pro Gln Glu Trp 3860 3865 3870 Lys Lys Lys Ser Leu Ile Gln Lys Leu Ile Leu Leu Arg Ala Met Arg 3875 3880 3885 Pro Asp Arg Met Thr Tyr Ala Leu Arg Asn Phe Val Glu Glu Lys Leu 3890 3895 3900 Gly Ala Lys Tyr Val Glu Arg Thr Arg Leu Asp Leu Val Lys Ala Phe 3905 3910 3915 3920 Glu Glu Ser Ser Pro Ala Thr Pro Ile Phe Phe Ile Leu Ser Pro Gly 3925 3930 3935 Val Asp Ala Leu Lys Asp Leu Glu Ile Leu Gly Lys Arg Leu Gly Phe 3940 3945 3950 Thr Ile Asp Ser Gly Lys Phe His Asn Val Ser Leu Gly Gln Gly Gln 3955 3960 3965 Glu Thr Val Ala Glu Val Ala Leu Glu Lys Ala Ser Lys Gly Gly His 3970 3975 3980 Trp Val Ile Leu Gln Asn Val His Leu Val Ala Lys Trp Leu Gly Thr 3985 3990 3995 4000 Leu Glu Lys Leu Leu Glu Arg Phe Ser Gln Gly Ser His Arg Asp Tyr 4005 4010 4015 Arg Val Phe Met Ser Ala Glu Ser Ala Pro Thr Pro Asp Glu His Ile 4020 4025 4030 Ile Pro Gln Gly Leu Leu Glu Asn Ser Ile Lys Ile Thr Asn Glu Pro 4035 4040 4045 Pro Thr Gly Met Leu Ala Asn Leu His Ala Ala Leu Tyr Asn Phe Asp 4050 4055 4060 Gln Asp Thr Leu Glu Ile Cys Ser Lys Glu Gln Glu Phe Lys Ser Ile 4065 4070 4075 4080 Leu Phe Ser Leu Cys Tyr Phe His Ala Cys Val Ala Gly Arg Leu Arg 4085 4090 4095 Phe Gly Pro Gln Gly Trp Ser Arg Ser Tyr Pro Phe Asn Pro Gly Asp 4100 4105 4110 Leu Thr Ile Cys Ala Ser Val Leu Tyr Asn Tyr Leu Glu Ala Asn Ser 4115 4120 4125 Lys Val Pro Trp Glu Asp Leu Arg Tyr Leu Phe Gly Glu Ile Met Tyr 4130 4135 4140 Gly Gly His Ile Thr Asp Asp Trp Asp Arg Lys Leu Cys Arg Val Tyr 4145 4150 4155 4160 Leu Glu Glu Phe Met Asn Pro Ser Leu Thr Glu Asp Glu Leu Met Leu 4165 4170 4175 Ala Pro Gly Phe Ala Ala Pro Pro Tyr Leu Asp Tyr Ala Gly Tyr His 4180 4185 4190 Gln Tyr Ile Glu Glu Met Leu Pro Pro Glu Ser Pro Ala Leu Tyr Gly 4195 4200 4205 Leu His Pro Asn Ala Glu Ile Glu Phe Leu Thr Val Thr Ser Asn Thr 4210 4215 4220 Leu Phe Arg Thr Leu Leu Glu Met Gln Pro Arg Asn Ala Leu Ser Gly 4225 4230 4235 4240 Asp Glu Leu Gly Gln Ser Thr Glu Glu Lys Val Lys Asn Val Leu Asp 4245 4250 4255 Asp Ile Leu Glu Lys Leu Pro Glu Glu Phe Asn Met Ala Glu Ile Met 4260 4265 4270 Gln Lys Asn Ser Asn Arg Ser Pro Tyr Val Leu Val Cys Phe Gln Glu 4275 4280 4285 Cys Glu Arg Met Asn Ile Leu Ile Arg Glu Ile Arg Ile Ser Leu Glu 4290 4295 4300 Gln Leu Asp Leu Ser Leu Lys Gly Glu Leu Ala Leu Ser Pro Ala Val 4305 4310 4315 4320 Glu Ala Gln Gln Phe Ala Leu Ser Tyr Asp Thr Val Pro Asp Thr Trp 4325 4330 4335 Ser Lys Leu Ala Tyr Pro Ser Thr Tyr Gly Leu Ala Gln Trp Phe Asn 4340 4345 4350 Asp Leu Leu Leu Arg Cys Arg Glu Leu Asp Thr Trp Thr Gln Asp Leu 4355 4360 4365 Thr Leu Pro Ala Val Val Trp Leu Ser Gly Phe Phe Asn Pro Gln Ser 4370 4375 4380 Phe Leu Thr Ala Ile Met Gln Thr Met Ala Arg Lys Asn Glu Trp Pro 4385 4390 4395 4400 Leu Asp Lys Thr Arg Leu Thr Ala Asp Val Thr Lys Lys Thr Lys Glu 4405 4410 4415 Asp Tyr Gly His Pro Pro Arg Glu Gly Ala Tyr Leu His Gly Leu Phe 4420 4425 4430 Met Glu Gly Ala Arg Trp Asp Thr Gln Ala Gly Thr Ile Val Glu Ala 4435 4440 4445 Arg Leu Lys Glu Leu Ala Cys Pro Met Pro Val Ile Phe Ala Lys Ala 4450 4455 4460 Thr Pro Val Asp Arg Gln Glu Thr Lys Gln Thr Tyr Glu Cys Pro Val 4465 4470 4475 4480 Tyr Arg Thr Lys Leu Arg Gly Pro Ser Tyr Ile Trp Thr Phe Arg Leu 4485 4490 4495 Lys Ser Glu Glu Lys Thr Ala Lys Trp Val Leu Ala Gly Val Ala Leu 4500 4505 4510 Leu Leu Glu Ala 4515 <210> 2 <211> 284 <212> PRT <213> Artificial Sequence <220> <223> NIPSNAP1 amino acid sequence <400> 2 Met Ala Pro Arg Leu Cys Ser Ile Ser Val Thr Ala Arg Arg Leu Leu 1 5 10 15 Gly Gly Pro Gly Pro Arg Ala Gly Asp Val Ala Ser Ala Ala Ala Ala 20 25 30 Arg Phe Tyr Ser Lys Asp Asn Glu Gly Ser Trp Phe Arg Ser Leu Phe 35 40 45 Val His Lys Val Asp Pro Arg Lys Asp Ala His Ser Thr Leu Leu Ser 50 55 60 Lys Lys Glu Thr Ser Asn Leu Tyr Lys Ile Gln Phe His Asn Val Lys 65 70 75 80 Pro Glu Tyr Leu Asp Ala Tyr Asn Ser Leu Thr Glu Ala Val Leu Pro 85 90 95 Lys Leu His Leu Asp Glu Asp Tyr Pro Cys Ser Leu Val Gly Asn Trp 100 105 110 Asn Thr Trp Tyr Gly Glu Gln Asp Gln Ala Val His Leu Trp Arg Phe 115 120 125 Ser Gly Gly Tyr Pro Ala Leu Met Asp Cys Met Asn Lys Leu Lys Asn 130 135 140 Asn Lys Glu Tyr Leu Glu Phe Arg Arg Glu Arg Ser Gln Met Leu Leu 145 150 155 160 Ser Arg Arg Asn Gln Leu Leu Leu Glu Phe Ser Phe Trp Asn Glu Pro 165 170 175 Gln Pro Arg Met Gly Pro Asn Ile Tyr Glu Leu Arg Thr Tyr Lys Leu 180 185 190 Lys Pro Gly Thr Met Ile Glu Trp Gly Asn Asn Trp Ala Arg Ala Ile 195 200 205 Lys Tyr Arg Gln Glu Asn Gln Glu Ala Val Gly Gly Phe Phe Ser Gln 210 215 220 Ile Gly Glu Leu Tyr Val Val His His Leu Trp Ala Tyr Lys Asp Leu 225 230 235 240 Gln Ser Arg Glu Glu Thr Arg Asn Ala Ala Trp Arg Lys Arg Gly Trp 245 250 255 Asp Glu Asn Val Tyr Tyr Thr Val Pro Leu Val Arg His Met Glu Ser 260 265 270 Arg Ile Met Ile Pro Leu Lys Ile Ser Pro Leu Gln 275 280 <210> 3 <211> 1338 <212> PRT <213> Artificial Sequence <220> <223> PFAS amino acid sequence <400> 3 Met Ser Pro Val Leu His Phe Tyr Val Arg Pro Ser Gly His Glu Gly 1 5 10 15 Ala Ala Pro Gly His Thr Arg Arg Lys Leu Gln Gly Lys Leu Pro Glu 20 25 30 Leu Gln Gly Val Glu Thr Glu Leu Cys Tyr Asn Val Asn Trp Thr Ala 35 40 45 Glu Ala Leu Pro Ser Ala Glu Glu Thr Lys Lys Leu Met Trp Leu Phe 50 55 60 Gly Cys Pro Leu Leu Leu Asp Asp Val Ala Arg Glu Ser Trp Leu Leu 65 70 75 80 Pro Gly Ser Asn Asp Leu Leu Leu Glu Val Gly Pro Arg Leu Asn Phe 85 90 95 Ser Thr Pro Thr Ser Thr Asn Ile Val Ser Val Cys Arg Ala Thr Gly 100 105 110 Leu Gly Pro Val Asp Arg Val Glu Thr Thr Arg Arg Tyr Arg Leu Ser 115 120 125 Phe Ala His Pro Pro Ser Ala Glu Val Glu Ala Ile Ala Leu Ala Thr 130 135 140 Leu His Asp Arg Met Thr Glu Gln His Phe Pro His Pro Ile Gln Ser 145 150 155 160 Phe Ser Pro Glu Ser Met Pro Glu Pro Leu Asn Gly Pro Ile Asn Ile 165 170 175 Leu Gly Glu Gly Arg Leu Ala Leu Glu Lys Ala Asn Gln Glu Leu Gly 180 185 190 Leu Ala Leu Asp Ser Trp Asp Leu Asp Phe Tyr Thr Lys Arg Phe Gln 195 200 205 Glu Leu Gln Arg Asn Pro Ser Thr Val Glu Ala Phe Asp Leu Ala Gln 210 215 220 Ser Asn Ser Glu His Ser Arg His Trp Phe Phe Lys Gly Gln Leu His 225 230 235 240 Val Asp Gly Gln Lys Leu Val His Ser Leu Phe Glu Ser Ile Met Ser 245 250 255 Thr Gln Glu Ser Ser Asn Pro Asn Asn Val Leu Lys Phe Cys Asp Asn 260 265 270 Ser Ser Ala Ile Gln Gly Lys Glu Val Arg Phe Leu Arg Pro Glu Asp 275 280 285 Pro Thr Arg Pro Ser Arg Phe Gln Gln Gln Gln Gly Leu Arg His Val 290 295 300 Val Phe Thr Ala Glu Thr His Asn Phe Pro Thr Gly Val Cys Pro Phe 305 310 315 320 Ser Gly Ala Thr Thr Gly Thr Gly Gly Arg Ile Arg Asp Val Gln Cys 325 330 335 Thr Gly Arg Gly Ala His Val Val Ala Gly Thr Ala Gly Tyr Cys Phe 340 345 350 Gly Asn Leu His Ile Pro Gly Tyr Asn Leu Pro Trp Glu Asp Pro Ser 355 360 365 Phe Gln Tyr Pro Gly Asn Phe Ala Arg Pro Leu Glu Val Ala Ile Glu 370 375 380 Ala Ser Asn Gly Ala Ser Asp Tyr Gly Asn Lys Phe Gly Glu Pro Val 385 390 395 400 Leu Ala Gly Phe Ala Arg Ser Leu Gly Leu Gln Leu Pro Asp Gly Gln 405 410 415 Arg Arg Glu Trp Ile Lys Pro Ile Met Phe Ser Gly Gly Ile Gly Ser 420 425 430 Met Glu Ala Asp His Ile Ser Lys Glu Ala Pro Glu Pro Gly Met Glu 435 440 445 Val Val Lys Val Gly Gly Pro Val Tyr Arg Ile Gly Val Gly Gly Gly 450 455 460 Ala Ala Ser Ser Val Gln Val Gln Gly Asp Asn Thr Ser Asp Leu Asp 465 470 475 480 Phe Gly Ala Val Gln Arg Gly Asp Pro Glu Met Glu Gln Lys Met Asn 485 490 495 Arg Val Ile Arg Ala Cys Val Glu Ala Pro Lys Gly Asn Pro Ile Cys 500 505 510 Ser Leu His Asp Gln Gly Ala Gly Gly Asn Gly Asn Val Leu Lys Glu 515 520 525 Leu Ser Asp Pro Ala Gly Ala Ile Ile Tyr Thr Ser Arg Phe Gln Leu 530 535 540 Gly Asp Pro Thr Leu Asn Ala Leu Glu Ile Trp Gly Ala Glu Tyr Gln 545 550 555 560 Glu Ser Asn Ala Leu Leu Leu Arg Ser Pro Asn Arg Asp Phe Leu Thr 565 570 575 His Val Ser Ala Arg Glu Arg Cys Pro Ala Cys Phe Val Gly Thr Ile 580 585 590 Thr Gly Asp Arg Arg Ile Val Leu Val Asp Asp Arg Glu Cys Pro Val 595 600 605 Arg Arg Asn Gly Gln Gly Asp Ala Pro Pro Thr Pro Leu Pro Thr Pro 610 615 620 Val Asp Leu Glu Leu Glu Trp Val Leu Gly Lys Met Pro Arg Lys Glu 625 630 635 640 Phe Phe Leu Gln Arg Lys Pro Pro Met Leu Gln Pro Leu Ala Leu Pro 645 650 655 Pro Gly Leu Ser Val His Gln Ala Leu Glu Arg Val Leu Arg Leu Pro 660 665 670 Ala Val Ala Ser Lys Arg Tyr Leu Thr Asn Lys Val Asp Arg Ser Val 675 680 685 Gly Gly Leu Val Ala Gln Gln Gln Cys Val Gly Pro Leu Gln Thr Pro 690 695 700 Leu Ala Asp Val Ala Val Val Ala Leu Ser His Glu Glu Leu Ile Gly 705 710 715 720 Ala Ala Thr Ala Leu Gly Glu Gln Pro Val Lys Ser Leu Leu Asp Pro 725 730 735 Lys Val Ala Ala Arg Leu Ala Val Ala Glu Ala Leu Thr Asn Leu Val 740 745 750 Phe Ala Leu Val Thr Asp Leu Arg Asp Val Lys Cys Ser Gly Asn Trp 755 760 765 Met Trp Ala Ala Lys Leu Pro Gly Glu Gly Ala Ala Leu Ala Asp Ala 770 775 780 Cys Glu Ala Met Val Ala Val Met Ala Ala Leu Gly Val Ala Val Asp 785 790 795 800 Gly Gly Lys Asp Ser Leu Ser Met Ala Ala Arg Val Gly Thr Glu Thr 805 810 815 Val Arg Ala Pro Gly Ser Leu Val Ile Ser Ala Tyr Ala Val Cys Pro 820 825 830 Asp Ile Thr Ala Thr Val Thr Pro Asp Leu Lys His Pro Glu Gly Arg 835 840 845 Gly His Leu Leu Tyr Val Ala Leu Ser Pro Gly Gln His Arg Leu Gly 850 855 860 Gly Thr Ala Leu Ala Gln Cys Phe Ser Gln Leu Gly Glu His Pro Pro 865 870 875 880 Asp Leu Asp Leu Pro Glu Asn Leu Val Arg Ala Phe Ser Ile Thr Gln 885 890 895 Gly Leu Leu Lys Asp Arg Leu Leu Cys Ser Gly His Asp Val Ser Asp 900 905 910 Gly Gly Leu Val Thr Cys Leu Leu Glu Met Ala Phe Ala Gly Asn Cys 915 920 925 Gly Leu Gln Val Asp Val Pro Val Pro Arg Val Asp Val Leu Ser Val 930 935 940 Leu Phe Ala Glu Glu Pro Gly Leu Val Leu Glu Val Gln Glu Pro Asp 945 950 955 960 Leu Ala Gln Val Leu Lys Arg Tyr Arg Asp Ala Gly Leu His Cys Leu 965 970 975 Glu Leu Gly His Thr Gly Glu Ala Gly Pro His Ala Met Val Arg Val 980 985 990 Ser Val Asn Gly Ala Val Val Leu Glu Glu Pro Val Gly Glu Leu Arg 995 1000 1005 Ala Leu Trp Glu Glu Thr Ser Phe Gln Leu Asp Arg Leu Gln Ala Glu 1010 1015 1020 Pro Arg Cys Val Ala Glu Glu Glu Arg Gly Leu Arg Glu Arg Met Gly 1025 1030 1035 1040 Pro Ser Tyr Cys Leu Pro Pro Thr Phe Pro Lys Ala Ser Val Pro Arg 1045 1050 1055 Glu Pro Gly Gly Pro Ser Pro Arg Val Ala Ile Leu Arg Glu Glu Gly 1060 1065 1070 Ser Asn Gly Asp Arg Glu Met Ala Asp Ala Phe His Leu Ala Gly Phe 1075 1080 1085 Glu Val Trp Asp Val Thr Met Gln Asp Leu Cys Ser Gly Ala Ile Gly 1090 1095 1100 Leu Asp Thr Phe Arg Gly Val Ala Phe Val Gly Gly Phe Ser Tyr Ala 1105 1110 1115 1120 Asp Val Leu Gly Ser Ala Lys Gly Trp Ala Ala Ala Val Thr Phe His 1125 1130 1135 Pro Arg Ala Gly Ala Glu Leu Arg Arg Phe Arg Lys Arg Pro Asp Thr 1140 1145 1150 Phe Ser Leu Gly Val Cys Asn Gly Cys Gln Leu Leu Ala Leu Leu Gly 1155 1160 1165 Trp Val Gly Gly Asp Pro Asn Glu Asp Ala Ala Glu Met Gly Pro Asp 1170 1175 1180 Ser Gln Pro Ala Arg Pro Gly Leu Leu Leu Arg His Asn Leu Ser Gly 1185 1190 1195 1200 Arg Tyr Glu Ser Arg Trp Ala Ser Val Arg Val Gly Pro Gly Pro Ala 1205 1210 1215 Leu Met Leu Arg Gly Met Glu Gly Ala Val Leu Pro Val Trp Ser Ala 1220 1225 1230 His Gly Glu Gly Tyr Val Ala Phe Ser Ser Pro Glu Leu Gln Ala Gln 1235 1240 1245 Ile Glu Ala Arg Gly Leu Ala Pro Leu His Trp Ala Asp Asp Asp Gly 1250 1255 1260 Asn Pro Thr Glu Gln Tyr Pro Leu Asn Pro Asn Gly Ser Pro Gly Gly 1265 1270 1275 1280 Val Ala Gly Ile Cys Ser Cys Asp Gly Arg His Leu Ala Val Met Pro 1285 1290 1295 His Pro Glu Arg Ala Val Arg Pro Trp Gln Trp Ala Trp Arg Pro Pro 1300 1305 1310 Pro Phe Asp Thr Leu Thr Thr Ser Pro Trp Leu Gln Leu Phe Ile Asn 1315 1320 1325 Ala Arg Asn Trp Thr Leu Glu Gly Ser Cys 1330 1335 <210> 4 <211> 833 <212> PRT <213> Artificial Sequence <220> <223> PPP4R3A amino acid sequence <400> 4 Met Thr Asp Thr Arg Arg Arg Val Lys Val Tyr Thr Leu Asn Glu Asp 1 5 10 15 Arg Gln Trp Asp Asp Arg Gly Thr Gly His Val Ser Ser Gly Tyr Val 20 25 30 Glu Arg Leu Lys Gly Met Ser Leu Leu Val Arg Ala Glu Ser Asp Gly 35 40 45 Ser Leu Leu Leu Glu Ser Lys Ile Asn Pro Asn Thr Ala Tyr Gln Lys 50 55 60 Gln Gln Asp Thr Leu Ile Val Trp Ser Glu Ala Glu Asn Tyr Asp Leu 65 70 75 80 Ala Leu Ser Phe Gln Glu Lys Ala Gly Cys Asp Glu Ile Trp Glu Lys 85 90 95 Ile Cys Gln Val Gln Gly Lys Asp Pro Ser Val Asp Ile Thr Gln Asp 100 105 110 Leu Val Asp Glu Ser Glu Glu Glu Arg Phe Asp Asp Met Ser Ser Pro 115 120 125 Gly Leu Glu Leu Pro Ser Cys Glu Leu Ser Arg Leu Glu Glu Ile Ala 130 135 140 Glu Leu Val Ala Ser Ser Leu Pro Ser Pro Leu Arg Arg Glu Lys Leu 145 150 155 160 Ala Leu Ala Leu Glu Asn Glu Gly Tyr Ile Lys Lys Leu Leu Glu Leu 165 170 175 Phe His Val Cys Glu Asp Leu Glu Asn Ile Glu Gly Leu His His Leu 180 185 190 Tyr Glu Ile Ile Lys Gly Ile Phe Leu Leu Asn Arg Thr Ala Leu Phe 195 200 205 Glu Val Met Phe Ser Glu Glu Cys Ile Met Asp Val Ile Gly Cys Leu 210 215 220 Glu Tyr Asp Pro Ala Leu Ser Gln Pro Arg Lys His Arg Glu Phe Leu 225 230 235 240 Thr Lys Thr Ala Lys Phe Lys Glu Val Ile Pro Ile Ser Asp Pro Glu 245 250 255 Leu Lys Gln Lys Ile His Gln Thr Tyr Arg Val Gln Tyr Ile Gln Asp 260 265 270 Met Val Leu Pro Thr Pro Ser Val Phe Glu Glu Asn Met Leu Ser Thr 275 280 285 Leu His Ser Phe Ile Phe Phe Asn Lys Val Glu Ile Val Gly Met Leu 290 295 300 Gln Glu Asp Glu Lys Phe Leu Thr Asp Leu Phe Ala Gln Leu Thr Asp 305 310 315 320 Glu Ala Thr Asp Glu Glu Lys Arg Gln Glu Leu Val Asn Phe Leu Lys 325 330 335 Glu Phe Cys Ala Phe Ser Gln Thr Leu Gln Pro Gln Asn Arg Asp Ala 340 345 350 Phe Phe Lys Thr Leu Ser Asn Met Gly Ile Leu Pro Ala Leu Glu Val 355 360 365 Ile Leu Gly Met Asp Asp Thr Gln Val Arg Ser Ala Ala Thr Asp Ile 370 375 380 Phe Ser Tyr Leu Val Glu Tyr Asn Pro Ser Met Val Arg Glu Phe Val 385 390 395 400 Met Gln Glu Ala Gln Gln Asn Asp Asp Val Ser Lys Lys Leu Thr Glu 405 410 415 Gln Lys Ile Thr Ser Lys Asp Ile Leu Leu Ile Asn Leu Ile Ile Glu 420 425 430 His Met Ile Cys Asp Thr Asp Pro Glu Leu Gly Gly Ala Val Gln Leu 435 440 445 Met Gly Leu Leu Arg Thr Leu Val Asp Pro Glu Asn Met Leu Ala Thr 450 455 460 Ala Asn Lys Thr Glu Lys Thr Glu Phe Leu Gly Phe Phe Tyr Lys His 465 470 475 480 Cys Met His Val Leu Thr Ala Pro Leu Leu Ala Asn Thr Thr Glu Asp 485 490 495 Lys Pro Ser Lys Asp Asp Phe Gln Thr Ala Gln Leu Leu Ala Leu Val 500 505 510 Leu Glu Leu Leu Thr Phe Cys Val Glu His His Thr Tyr His Ile Lys 515 520 525 Asn Tyr Ile Ile Asn Lys Asp Ile Leu Arg Arg Val Leu Val Leu Met 530 535 540 Ala Ser Lys His Ala Phe Leu Ala Leu Cys Ala Leu Arg Phe Lys Arg 545 550 555 560 Lys Ile Ile Gly Leu Lys Asp Glu Phe Tyr Asn Arg Tyr Ile Met Lys 565 570 575 Ser Phe Leu Phe Glu Pro Val Val Lys Ala Phe Leu Asn Asn Gly Ser 580 585 590 Arg Tyr Asn Leu Met Asn Ser Ala Ile Ile Glu Met Phe Glu Phe Ile 595 600 605 Arg Val Glu Asp Ile Lys Ser Leu Thr Ala His Val Ile Glu Asn Tyr 610 615 620 Trp Lys Ala Leu Glu Asp Val Asp Tyr Val Gln Thr Phe Lys Gly Leu 625 630 635 640 Lys Leu Arg Phe Glu Gln Gln Arg Glu Arg Gln Asp Asn Pro Lys Leu 645 650 655 Asp Ser Met Arg Ser Ile Leu Arg Asn His Arg Tyr Arg Arg Asp Ala 660 665 670 Arg Thr Leu Glu Asp Glu Glu Glu Met Trp Phe Asn Thr Asp Glu Asp 675 680 685 Asp Met Glu Asp Gly Glu Ala Val Val Ser Pro Ser Asp Lys Thr Lys 690 695 700 Asn Asp Asp Asp Ile Met Asp Pro Ile Ser Lys Phe Met Glu Arg Lys 705 710 715 720 Lys Leu Lys Glu Ser Glu Glu Lys Glu Val Leu Leu Lys Thr Asn Leu 725 730 735 Ser Gly Arg Gln Ser Pro Ser Phe Lys Leu Ser Leu Ser Ser Gly Thr 740 745 750 Lys Thr Asn Leu Thr Ser Gln Ser Ser Thr Thr Asn Leu Pro Gly Ser 755 760 765 Pro Gly Ser Pro Gly Ser Pro Gly Ser Pro Gly Ser Pro Gly Ser Val 770 775 780 Pro Lys Asn Thr Ser Gln Thr Ala Ala Ile Thr Thr Lys Gly Gly Leu 785 790 795 800 Val Gly Leu Val Asp Tyr Pro Asp Asp Asp Glu Asp Asp Asp Glu Asp 805 810 815 Glu Asp Lys Glu Asp Thr Leu Pro Leu Ser Lys Lys Ala Lys Phe Asp 820 825 830 Ser <210> 5 <211> 146 <212> PRT <213> Artificial Sequence <220> <223> MCFD2 amino acid sequence <400> 5 Met Thr Met Arg Ser Leu Leu Arg Thr Pro Phe Leu Cys Gly Leu Leu 1 5 10 15 Trp Ala Phe Cys Ala Pro Gly Ala Arg Ala Glu Glu Pro Ala Ala Ser 20 25 30 Phe Ser Gln Pro Gly Ser Met Gly Leu Asp Lys Asn Thr Val His Asp 35 40 45 Gln Glu His Ile Met Glu His Leu Glu Gly Val Ile Asn Lys Pro Glu 50 55 60 Ala Glu Met Ser Pro Gln Glu Leu Gln Leu His Tyr Phe Lys Met His 65 70 75 80 Asp Tyr Asp Gly Asn Asn Leu Leu Asp Gly Leu Glu Leu Ser Thr Ala 85 90 95 Ile Thr His Val His Lys Glu Glu Gly Ser Glu Gln Ala Pro Leu Met 100 105 110 Ser Glu Asp Glu Leu Ile Asn Ile Ile Asp Gly Val Leu Arg Asp Asp 115 120 125 Asp Lys Asn Asn Asp Gly Tyr Ile Asp Tyr Ala Glu Phe Ala Lys Ser 130 135 140 Leu Gln 145 <210> 6 <211> 1151 <212> PRT <213> Artificial Sequence <220> <223> NRDC amino acid sequence <400> 6 Met Leu Arg Arg Val Thr Val Ala Ala Val Cys Ala Thr Arg Arg Lys 1 5 10 15 Leu Cys Glu Ala Gly Arg Glu Leu Ala Ala Leu Trp Gly Ile Glu Thr 20 25 30 Arg Gly Arg Cys Glu Asp Ser Ala Ala Ala Arg Pro Phe Pro Ile Leu 35 40 45 Ala Met Pro Gly Arg Asn Lys Ala Lys Ser Thr Cys Ser Cys Pro Asp 50 55 60 Leu Gln Pro Asn Gly Gln Asp Leu Gly Glu Asn Ser Arg Val Ala Arg 65 70 75 80 Leu Gly Ala Asp Glu Ser Glu Glu Glu Gly Arg Arg Gly Ser Leu Ser 85 90 95 Asn Ala Gly Asp Pro Glu Ile Val Lys Ser Pro Ser Asp Pro Lys Gln 100 105 110 Tyr Arg Tyr Ile Lys Leu Gln Asn Gly Leu Gln Ala Leu Leu Ile Ser 115 120 125 Asp Leu Ser Asn Met Glu Gly Lys Thr Gly Asn Thr Thr Asp Asp Glu 130 135 140 Glu Glu Glu Glu Val Glu Glu Glu Glu Glu Asp Asp Asp Glu Asp Ser 145 150 155 160 Gly Ala Glu Ile Glu Asp Asp Asp Glu Glu Gly Phe Asp Asp Glu Asp 165 170 175 Glu Phe Asp Asp Glu His Asp Asp Asp Leu Asp Thr Glu Asp Asn Glu 180 185 190 Leu Glu Glu Leu Glu Glu Arg Ala Glu Ala Arg Lys Lys Thr Thr Glu 195 200 205 Lys Gln Ser Ala Ala Ala Leu Cys Val Gly Val Gly Ser Phe Ala Asp 210 215 220 Pro Asp Asp Leu Pro Gly Leu Ala His Phe Leu Glu His Met Val Phe 225 230 235 240 Met Gly Ser Leu Lys Tyr Pro Asp Glu Asn Gly Phe Asp Ala Phe Leu 245 250 255 Lys Lys His Gly Gly Ser Asp Asn Ala Ser Thr Asp Cys Glu Arg Thr 260 265 270 Val Phe Gln Phe Asp Val Gln Arg Lys Tyr Phe Lys Glu Ala Leu Asp 275 280 285 Arg Trp Ala Gln Phe Phe Ile His Pro Leu Met Ile Arg Asp Ala Ile 290 295 300 Asp Arg Glu Val Glu Ala Val Asp Ser Glu Tyr Gln Leu Ala Arg Pro 305 310 315 320 Ser Asp Ala Asn Arg Lys Glu Met Leu Phe Gly Ser Leu Ala Arg Pro 325 330 335 Gly His Pro Met Gly Lys Phe Phe Trp Gly Asn Ala Glu Thr Leu Lys 340 345 350 His Glu Pro Arg Lys Asn Asn Ile Asp Thr His Ala Arg Leu Arg Glu 355 360 365 Phe Trp Met Arg Tyr Tyr Ser Ser His Tyr Met Thr Leu Val Val Gln 370 375 380 Ser Lys Glu Thr Leu Asp Thr Leu Glu Lys Trp Val Thr Glu Ile Phe 385 390 395 400 Ser Gln Ile Pro Asn Asn Gly Leu Pro Arg Pro Asn Phe Gly His Leu 405 410 415 Thr Asp Pro Phe Asp Thr Pro Ala Phe Asn Lys Leu Tyr Arg Val Val 420 425 430 Pro Ile Arg Lys Ile His Ala Leu Thr Ile Thr Trp Ala Leu Pro Pro 435 440 445 Gln Gln Gln His Tyr Arg Val Lys Pro Leu His Tyr Ile Ser Trp Leu 450 455 460 Val Gly His Glu Gly Lys Gly Ser Ile Leu Ser Phe Leu Arg Lys Lys 465 470 475 480 Cys Trp Ala Leu Ala Leu Phe Gly Gly Asn Gly Glu Thr Gly Phe Glu 485 490 495 Gln Asn Ser Thr Tyr Ser Val Phe Ser Ile Ser Ile Thr Leu Thr Asp 500 505 510 Glu Gly Tyr Glu His Phe Tyr Glu Val Ala Tyr Thr Val Phe Gln Tyr 515 520 525 Leu Lys Met Leu Gln Lys Leu Gly Pro Glu Lys Arg Ile Phe Glu Glu 530 535 540 Ile Arg Lys Ile Glu Asp Asn Glu Phe His Tyr Gln Glu Gln Thr Asp 545 550 555 560 Pro Val Glu Tyr Val Glu Asn Met Cys Glu Asn Met Gln Leu Tyr Pro 565 570 575 Leu Gln Asp Ile Leu Thr Gly Asp Gln Leu Leu Phe Glu Tyr Lys Pro 580 585 590 Glu Val Ile Gly Glu Ala Leu Asn Gln Leu Val Pro Gln Lys Ala Asn 595 600 605 Leu Val Leu Leu Ser Gly Ala Asn Glu Gly Lys Cys Asp Leu Lys Glu 610 615 620 Lys Trp Phe Gly Thr Gln Tyr Ser Ile Glu Asp Ile Glu Asn Ser Trp 625 630 635 640 Ala Glu Leu Trp Asn Ser Asn Phe Glu Leu Asn Pro Asp Leu His Leu 645 650 655 Pro Ala Glu Asn Lys Tyr Ile Ala Thr Asp Phe Thr Leu Lys Ala Phe 660 665 670 Asp Cys Pro Glu Thr Glu Tyr Pro Val Lys Ile Val Asn Thr Pro Gln 675 680 685 Gly Cys Leu Trp Tyr Lys Lys Asp Asn Lys Phe Lys Ile Pro Lys Ala 690 695 700 Tyr Ile Arg Phe His Leu Ile Ser Pro Leu Ile Gln Lys Ser Ala Ala 705 710 715 720 Asn Val Val Leu Phe Asp Ile Phe Val Asn Ile Leu Thr His Asn Leu 725 730 735 Ala Glu Pro Ala Tyr Glu Ala Asp Val Ala Gln Leu Glu Tyr Lys Leu 740 745 750 Val Ala Gly Glu His Gly Leu Ile Ile Arg Val Lys Gly Phe Asn His 755 760 765 Lys Leu Pro Leu Leu Phe Gln Leu Ile Ile Asp Tyr Leu Ala Glu Phe 770 775 780 Asn Ser Thr Pro Ala Val Phe Thr Met Ile Thr Glu Gln Leu Lys Lys 785 790 795 800 Thr Tyr Phe Asn Ile Leu Ile Lys Pro Glu Thr Leu Ala Lys Asp Val 805 810 815 Arg Leu Leu Ile Leu Glu Tyr Ala Arg Trp Ser Met Ile Asp Lys Tyr 820 825 830 Gln Ala Leu Met Asp Gly Leu Ser Leu Glu Ser Leu Leu Ser Phe Val 835 840 845 Lys Glu Phe Lys Ser Gln Leu Phe Val Glu Gly Leu Val Gln Gly Asn 850 855 860 Val Thr Ser Thr Glu Ser Met Asp Phe Leu Lys Tyr Val Val Asp Lys 865 870 875 880 Leu Asn Phe Lys Pro Leu Glu Gln Glu Met Pro Val Gln Phe Gln Val 885 890 895 Val Glu Leu Pro Ser Gly His His Leu Cys Lys Val Lys Ala Leu Asn 900 905 910 Lys Gly Asp Ala Asn Ser Glu Val Thr Val Tyr Tyr Gln Ser Gly Thr 915 920 925 Arg Ser Leu Arg Glu Tyr Thr Leu Met Glu Leu Leu Val Met His Met 930 935 940 Glu Glu Pro Cys Phe Asp Phe Leu Arg Thr Lys Gln Thr Leu Gly Tyr 945 950 955 960 His Val Tyr Pro Thr Cys Arg Asn Thr Ser Gly Ile Leu Gly Phe Ser 965 970 975 Val Thr Val Gly Thr Gln Ala Thr Lys Tyr Asn Ser Glu Val Val Asp 980 985 990 Lys Lys Ile Glu Glu Phe Leu Ser Ser Phe Glu Glu Lys Ile Glu Asn 995 1000 1005 Leu Thr Glu Glu Ala Phe Asn Thr Gln Val Thr Ala Leu Ile Lys Leu 1010 1015 1020 Lys Glu Cys Glu Asp Thr His Leu Gly Glu Glu Val Asp Arg Asn Trp 1025 1030 1035 1040 Asn Glu Val Val Thr Gln Gln Tyr Leu Phe Asp Arg Leu Ala His Glu 1045 1050 1055 Ile Glu Ala Leu Lys Ser Phe Ser Lys Ser Asp Leu Val Asn Trp Phe 1060 1065 1070 Lys Ala His Arg Gly Pro Gly Ser Lys Met Leu Ser Val His Val Val 1075 1080 1085 Gly Tyr Gly Lys Tyr Glu Leu Glu Glu Asp Gly Thr Pro Ser Ser Glu 1090 1095 1100 Asp Ser Asn Ser Ser Cys Glu Val Met Gln Leu Thr Tyr Leu Pro Thr 1105 1110 1115 1120 Ser Pro Leu Leu Ala Asp Cys Ile Ile Pro Ile Thr Asp Ile Arg Ala 1125 1130 1135 Phe Thr Thr Thr Leu Asn Leu Leu Pro Tyr His Lys Ile Val Lys 1140 1145 1150 <210> 7 <211> 257 <212> PRT <213> Artificial Sequence <220> <223> ALYREF amino acid sequence <400> 7 Met Ala Asp Lys Met Asp Met Ser Leu Asp Asp Ile Ile Lys Leu Asn 1 5 10 15 Arg Ser Gln Arg Gly Gly Arg Gly Gly Gly Arg Gly Arg Gly Arg Ala 20 25 30 Gly Ser Gln Gly Gly Arg Gly Gly Gly Ala Gln Ala Ala Ala Arg Val 35 40 45 Asn Arg Gly Gly Gly Pro Ile Arg Asn Arg Pro Ala Ile Ala Arg Gly 50 55 60 Ala Ala Gly Gly Gly Gly Arg Asn Arg Pro Ala Pro Tyr Ser Arg Pro 65 70 75 80 Lys Gln Leu Pro Asp Lys Trp Gln His Asp Leu Phe Asp Ser Gly Phe 85 90 95 Gly Gly Gly Ala Gly Val Glu Thr Gly Gly Lys Leu Leu Val Ser Asn 100 105 110 Leu Asp Phe Gly Val Ser Asp Ala Asp Ile Gln Glu Leu Phe Ala Glu 115 120 125 Phe Gly Thr Leu Lys Lys Ala Ala Val His Tyr Asp Arg Ser Gly Arg 130 135 140 Ser Leu Gly Thr Ala Asp Val His Phe Glu Arg Lys Ala Asp Ala Leu 145 150 155 160 Lys Ala Met Lys Gln Tyr Asn Gly Val Pro Leu Asp Gly Arg Pro Met 165 170 175 Asn Ile Gln Leu Val Thr Ser Gln Ile Asp Ala Gln Arg Arg Pro Ala 180 185 190 Gln Ser Val Asn Arg Gly Gly Met Thr Arg Asn Arg Gly Ala Gly Gly 195 200 205 Phe Gly Gly Gly Gly Gly Thr Arg Arg Gly Thr Arg Gly Gly Ala Arg 210 215 220 Gly Arg Gly Arg Gly Ala Gly Arg Asn Ser Lys Gln Gln Leu Ser Ala 225 230 235 240 Glu Glu Leu Asp Ala Gln Leu Asp Ala Tyr Asn Ala Arg Met Asp Thr 245 250 255 Ser <210> 8 <211> 736 <212> PRT <213> Artificial Sequence <220> <223> HSD17B4 amino acid sequence <400> 8 Met Gly Ser Pro Leu Arg Phe Asp Gly Arg Val Val Leu Val Thr Gly 1 5 10 15 Ala Gly Ala Gly Leu Gly Arg Ala Tyr Ala Leu Ala Phe Ala Glu Arg 20 25 30 Gly Ala Leu Val Val Val Asn Asp Leu Gly Gly Asp Phe Lys Gly Val 35 40 45 Gly Lys Gly Ser Leu Ala Ala Asp Lys Val Val Glu Glu Ile Arg Arg 50 55 60 Arg Gly Gly Lys Ala Val Ala Asn Tyr Asp Ser Val Glu Glu Gly Glu 65 70 75 80 Lys Val Val Lys Thr Ala Leu Asp Ala Phe Gly Arg Ile Asp Val Val 85 90 95 Val Asn Asn Ala Gly Ile Leu Arg Asp Arg Ser Phe Ala Arg Ile Ser 100 105 110 Asp Glu Asp Trp Asp Ile Ile His Arg Val His Leu Arg Gly Ser Phe 115 120 125 Gln Val Thr Arg Ala Ala Trp Glu His Met Lys Lys Gln Lys Tyr Gly 130 135 140 Arg Ile Ile Met Thr Ser Ser Ala Ser Gly Ile Tyr Gly Asn Phe Gly 145 150 155 160 Gln Ala Asn Tyr Ser Ala Ala Lys Leu Gly Leu Leu Gly Leu Ala Asn 165 170 175 Ser Leu Ala Ile Glu Gly Arg Lys Ser Asn Ile His Cys Asn Thr Ile 180 185 190 Ala Pro Asn Ala Gly Ser Arg Met Thr Gln Thr Val Met Pro Glu Asp 195 200 205 Leu Val Glu Ala Leu Lys Pro Glu Tyr Val Ala Pro Leu Val Leu Trp 210 215 220 Leu Cys His Glu Ser Cys Glu Glu Asn Gly Gly Leu Phe Glu Val Gly 225 230 235 240 Ala Gly Trp Ile Gly Lys Leu Arg Trp Glu Arg Thr Leu Gly Ala Ile 245 250 255 Val Arg Gln Lys Asn His Pro Met Thr Pro Glu Ala Val Lys Ala Asn 260 265 270 Trp Lys Lys Ile Cys Asp Phe Glu Asn Ala Ser Lys Pro Gln Ser Ile 275 280 285 Gln Glu Ser Thr Gly Ser Ile Ile Glu Val Leu Ser Lys Ile Asp Ser 290 295 300 Glu Gly Gly Val Ser Ala Asn His Thr Ser Arg Ala Thr Ser Thr Ala 305 310 315 320 Thr Ser Gly Phe Ala Gly Ala Ile Gly Gln Lys Leu Pro Pro Phe Ser 325 330 335 Tyr Ala Tyr Thr Glu Leu Glu Ala Ile Met Tyr Ala Leu Gly Val Gly 340 345 350 Ala Ser Ile Lys Asp Pro Lys Asp Leu Lys Phe Ile Tyr Glu Gly Ser 355 360 365 Ser Asp Phe Ser Cys Leu Pro Thr Phe Gly Val Ile Ile Gly Gln Lys 370 375 380 Ser Met Met Gly Gly Gly Leu Ala Glu Ile Pro Gly Leu Ser Ile Asn 385 390 395 400 Phe Ala Lys Val Leu His Gly Glu Gln Tyr Leu Glu Leu Tyr Lys Pro 405 410 415 Leu Pro Arg Ala Gly Lys Leu Lys Cys Glu Ala Val Val Ala Asp Val 420 425 430 Leu Asp Lys Gly Ser Gly Val Val Ile Ile Met Asp Val Tyr Ser Tyr 435 440 445 Ser Glu Lys Glu Leu Ile Cys His Asn Gln Phe Ser Leu Phe Leu Val 450 455 460 Gly Ser Gly Gly Phe Gly Gly Lys Arg Thr Ser Asp Lys Val Lys Val 465 470 475 480 Ala Val Ala Ile Pro Asn Arg Pro Pro Asp Ala Val Leu Thr Asp Thr 485 490 495 Thr Ser Leu Asn Gln Ala Ala Leu Tyr Arg Leu Ser Gly Asp Trp Asn 500 505 510 Pro Leu His Ile Asp Pro Asn Phe Ala Ser Leu Ala Gly Phe Asp Lys 515 520 525 Pro Ile Leu His Gly Leu Cys Thr Phe Gly Phe Ser Ala Arg Arg Val 530 535 540 Leu Gln Gln Phe Ala Asp Asn Asp Val Ser Arg Phe Lys Ala Ile Lys 545 550 555 560 Ala Arg Phe Ala Lys Pro Val Tyr Pro Gly Gln Thr Leu Gln Thr Glu 565 570 575 Met Trp Lys Glu Gly Asn Arg Ile His Phe Gln Thr Lys Val Gln Glu 580 585 590 Thr Gly Asp Ile Val Ile Ser Asn Ala Tyr Val Asp Leu Ala Pro Thr 595 600 605 Ser Gly Thr Ser Ala Lys Thr Pro Ser Glu Gly Gly Lys Leu Gln Ser 610 615 620 Thr Phe Val Phe Glu Glu Ile Gly Arg Arg Leu Lys Asp Ile Gly Pro 625 630 635 640 Glu Val Val Lys Lys Val Asn Ala Val Phe Glu Trp His Ile Thr Lys 645 650 655 Gly Gly Asn Ile Gly Ala Lys Trp Thr Ile Asp Leu Lys Ser Gly Ser 660 665 670 Gly Lys Val Tyr Gln Gly Pro Ala Lys Gly Ala Ala Asp Thr Thr Ile 675 680 685 Ile Leu Ser Asp Glu Asp Phe Met Glu Val Val Leu Gly Lys Leu Asp 690 695 700 Pro Gln Lys Ala Phe Phe Ser Gly Arg Leu Lys Ala Arg Gly Asn Ile 705 710 715 720 Met Leu Ser Gln Lys Leu Gln Met Ile Leu Lys Asp Tyr Ala Lys Leu 725 730 735 <210> 9 <211> 629 <212> PRT <213> Artificial Sequence <220> <223> PLS1 amino acid sequence <400> 9 Met Glu Asn Ser Thr Thr Thr Ile Ser Arg Glu Glu Leu Glu Glu Leu 1 5 10 15 Gln Glu Ala Phe Asn Lys Ile Asp Ile Asp Asn Ser Gly Tyr Val Ser 20 25 30 Asp Tyr Glu Leu Gln Asp Leu Phe Lys Glu Ala Ser Leu Pro Leu Pro 35 40 45 Gly Tyr Lys Val Arg Glu Ile Val Glu Lys Ile Leu Ser Val Ala Asp 50 55 60 Ser Asn Lys Asp Gly Lys Ile Ser Phe Glu Glu Phe Val Ser Leu Met 65 70 75 80 Gln Glu Leu Lys Ser Lys Asp Ile Ser Lys Thr Phe Arg Lys Ile Ile 85 90 95 Asn Lys Arg Glu Gly Ile Thr Ala Ile Gly Gly Thr Ser Thr Ile Ser 100 105 110 Ser Glu Gly Thr Gln His Ser Tyr Ser Glu Glu Glu Lys Val Ala Phe 115 120 125 Val Asn Trp Ile Asn Lys Ala Leu Glu Asn Asp Pro Asp Cys Lys His 130 135 140 Leu Ile Pro Met Asn Pro Asn Asp Asp Ser Leu Phe Lys Ser Leu Ala 145 150 155 160 Asp Gly Ile Leu Leu Cys Lys Met Ile Asn Leu Ser Glu Pro Asp Thr 165 170 175 Ile Asp Glu Arg Ala Ile Asn Lys Lys Lys Leu Thr Pro Phe Thr Ile 180 185 190 Ser Glu Asn Leu Asn Leu Ala Leu Asn Ser Ala Ser Ala Ile Gly Cys 195 200 205 Thr Val Val Asn Ile Gly Ala Ser Asp Leu Lys Glu Gly Lys Pro His 210 215 220 Leu Val Leu Gly Leu Leu Trp Gln Ile Ile Lys Val Gly Leu Phe Ala 225 230 235 240 Asp Ile Glu Ile Ser Arg Asn Glu Ala Leu Ile Ala Leu Leu Asn Glu 245 250 255 Gly Glu Glu Leu Glu Glu Leu Met Lys Leu Ser Pro Glu Glu Leu Leu 260 265 270 Leu Arg Trp Val Asn Tyr His Leu Thr Asn Ala Gly Trp His Thr Ile 275 280 285 Ser Asn Phe Ser Gln Asp Ile Lys Asp Ser Arg Ala Tyr Phe His Leu 290 295 300 Leu Asn Gln Ile Ala Pro Lys Gly Gly Glu Asp Gly Pro Ala Ile Ala 305 310 315 320 Ile Asp Leu Ser Gly Ile Asn Glu Thr Asn Asp Leu Lys Arg Ala Gly 325 330 335 Leu Met Leu Gln Glu Ala Asp Lys Leu Gly Cys Lys Gln Phe Val Thr 340 345 350 Pro Ala Asp Val Val Ser Gly Asn Pro Lys Leu Asn Leu Ala Phe Val 355 360 365 Ala Asn Leu Phe Asn Thr Tyr Pro Cys Leu His Lys Pro Asn Asn Asn 370 375 380 Asp Ile Asp Met Asn Leu Leu Glu Gly Glu Ser Lys Glu Glu Arg Thr 385 390 395 400 Phe Arg Asn Trp Met Asn Ser Leu Gly Val Asn Pro Tyr Ile Asn His 405 410 415 Leu Tyr Ser Asp Leu Ala Asp Ala Leu Val Ile Phe Gln Leu Tyr Glu 420 425 430 Met Ile Arg Val Pro Val Asn Trp Ser His Val Asn Lys Pro Pro Tyr 435 440 445 Pro Ala Leu Gly Gly Asn Met Lys Lys Ile Glu Asn Cys Asn Tyr Ala 450 455 460 Val Glu Leu Gly Lys Asn Lys Ala Lys Phe Ser Leu Val Gly Ile Ala 465 470 475 480 Gly Gln Asp Leu Asn Glu Gly Asn Ser Thr Leu Thr Leu Ala Leu Val 485 490 495 Trp Gln Leu Met Arg Arg Tyr Thr Leu Asn Val Leu Ser Asp Leu Gly 500 505 510 Glu Gly Glu Lys Val Asn Asp Glu Ile Ile Ile Lys Trp Val Asn Gln 515 520 525 Thr Leu Lys Ser Ala Asn Lys Lys Thr Ser Ile Ser Ser Phe Lys Asp 530 535 540 Lys Ser Ile Ser Thr Ser Leu Pro Val Leu Asp Leu Ile Asp Ala Ile 545 550 555 560 Ala Pro Asn Ala Val Arg Gln Glu Met Ile Arg Arg Glu Asn Leu Ser 565 570 575 Asp Glu Asp Lys Leu Asn Asn Ala Lys Tyr Ala Ile Ser Val Ala Arg 580 585 590 Lys Ile Gly Ala Arg Ile Tyr Ala Leu Pro Asp Asp Leu Val Glu Val 595 600 605 Lys Pro Lys Met Val Met Thr Val Phe Ala Cys Leu Met Gly Lys Gly 610 615 620 Leu Asn Arg Ile Lys 625 <210> 10 <211> 388 <212> PRT <213> Artificial Sequence <220> <223> PGC amino acid sequence <400> 10 Met Lys Trp Met Val Val Val Leu Val Cys Leu Gln Leu Leu Glu Ala 1 5 10 15 Ala Val Val Lys Val Pro Leu Lys Lys Phe Lys Ser Ile Arg Glu Thr 20 25 30 Met Lys Glu Lys Gly Leu Leu Gly Glu Phe Leu Arg Thr His Lys Tyr 35 40 45 Asp Pro Ala Trp Lys Tyr Arg Phe Gly Asp Leu Ser Val Thr Tyr Glu 50 55 60 Pro Met Ala Tyr Met Asp Ala Ala Tyr Phe Gly Glu Ile Ser Ile Gly 65 70 75 80 Thr Pro Pro Gln Asn Phe Leu Val Leu Phe Asp Thr Gly Ser Ser Asn 85 90 95 Leu Trp Val Pro Ser Val Tyr Cys Gln Ser Gln Ala Cys Thr Ser His 100 105 110 Ser Arg Phe Asn Pro Ser Glu Ser Ser Thr Tyr Ser Thr Asn Gly Gln 115 120 125 Thr Phe Ser Leu Gln Tyr Gly Ser Gly Ser Leu Thr Gly Phe Phe Gly 130 135 140 Tyr Asp Thr Leu Thr Val Gln Ser Ile Gln Val Pro Asn Gln Glu Phe 145 150 155 160 Gly Leu Ser Glu Asn Glu Pro Gly Thr Asn Phe Val Tyr Ala Gln Phe 165 170 175 Asp Gly Ile Met Gly Leu Ala Tyr Pro Ala Leu Ser Val Asp Glu Ala 180 185 190 Thr Thr Ala Met Gln Gly Met Val Gln Glu Gly Ala Leu Thr Ser Pro 195 200 205 Val Phe Ser Val Tyr Leu Ser Asn Gln Gln Gly Ser Ser Gly Gly Ala 210 215 220 Val Val Phe Gly Gly Val Asp Ser Ser Leu Tyr Thr Gly Gln Ile Tyr 225 230 235 240 Trp Ala Pro Val Thr Gln Glu Leu Tyr Trp Gln Ile Gly Ile Glu Glu 245 250 255 Phe Leu Ile Gly Gly Gln Ala Ser Gly Trp Cys Ser Glu Gly Cys Gln 260 265 270 Ala Ile Val Asp Thr Gly Thr Ser Leu Leu Thr Val Pro Gln Gln Tyr 275 280 285 Met Ser Ala Leu Leu Gln Ala Thr Gly Ala Gln Glu Asp Glu Tyr Gly 290 295 300 Gln Phe Leu Val Asn Cys Asn Ser Ile Gln Asn Leu Pro Ser Leu Thr 305 310 315 320 Phe Ile Ile Asn Gly Val Glu Phe Pro Leu Pro Pro Ser Ser Tyr Ile 325 330 335 Leu Ser Asn Asn Gly Tyr Cys Thr Val Gly Val Glu Pro Thr Tyr Leu 340 345 350 Ser Ser Gln Asn Gly Gln Pro Leu Trp Ile Leu Gly Asp Val Phe Leu 355 360 365 Arg Ser Tyr Tyr Ser Val Tyr Asp Leu Gly Asn Asn Arg Val Gly Phe 370 375 380 Ala Thr Ala Ala 385 <210> 11 <211> 441 <212> PRT <213> Artificial Sequence <220> <223> ARMT1 amino acid sequence <400> 11 Met Ala Val Val Pro Ala Ser Leu Ser Gly Gln Asp Val Gly Ser Phe 1 5 10 15 Ala Tyr Leu Thr Ile Lys Asp Arg Ile Pro Gln Ile Leu Thr Lys Val 20 25 30 Ile Asp Thr Leu His Arg His Lys Ser Glu Phe Phe Glu Lys His Gly 35 40 45 Glu Glu Gly Val Glu Ala Glu Lys Lys Ala Ile Ser Leu Leu Ser Lys 50 55 60 Leu Arg Asn Glu Leu Gln Thr Asp Lys Pro Phe Ile Pro Leu Val Glu 65 70 75 80 Lys Phe Val Asp Thr Asp Ile Trp Asn Gln Tyr Leu Glu Tyr Gln Gln 85 90 95 Ser Leu Leu Asn Glu Ser Asp Gly Lys Ser Arg Trp Phe Tyr Ser Pro 100 105 110 Trp Leu Leu Val Glu Cys Tyr Met Tyr Arg Arg Ile His Glu Ala Ile 115 120 125 Ile Gln Ser Pro Pro Ile Asp Tyr Phe Asp Val Phe Lys Glu Ser Lys 130 135 140 Glu Gln Asn Phe Tyr Gly Ser Gln Glu Ser Ile Ile Ala Leu Cys Thr 145 150 155 160 His Leu Gln Gln Leu Ile Arg Thr Ile Glu Asp Leu Asp Glu Asn Gln 165 170 175 Leu Lys Asp Glu Phe Phe Lys Leu Leu Gln Ile Ser Leu Trp Gly Asn 180 185 190 Lys Cys Asp Leu Ser Leu Ser Gly Gly Glu Ser Ser Ser Gln Asn Thr 195 200 205 Asn Val Leu Asn Ser Leu Glu Asp Leu Lys Pro Phe Ile Leu Leu Asn 210 215 220 Asp Met Glu His Leu Trp Ser Leu Leu Ser Asn Cys Lys Lys Thr Arg 225 230 235 240 Glu Lys Ala Ser Ala Thr Arg Val Tyr Ile Val Leu Asp Asn Ser Gly 245 250 255 Phe Glu Leu Val Thr Asp Leu Ile Leu Ala Asp Phe Leu Leu Ser Ser 260 265 270 Glu Leu Ala Thr Glu Val His Phe Tyr Gly Lys Thr Ile Pro Trp Phe 275 280 285 Val Ser Asp Thr Thr Ile His Asp Phe Asn Trp Leu Ile Glu Gln Val 290 295 300 Lys His Ser Asn His Lys Trp Met Ser Lys Cys Gly Ala Asp Trp Glu 305 310 315 320 Glu Tyr Ile Lys Met Gly Lys Trp Val Tyr His Asn His Ile Phe Trp 325 330 335 Thr Leu Pro His Glu Tyr Cys Ala Met Pro Gln Val Ala Pro Asp Leu 340 345 350 Tyr Ala Glu Leu Gln Lys Ala His Leu Ile Leu Phe Lys Gly Asp Leu 355 360 365 Asn Tyr Arg Lys Leu Thr Gly Asp Arg Lys Trp Glu Phe Ser Val Pro 370 375 380 Phe His Gln Ala Leu Asn Gly Phe His Pro Ala Pro Leu Cys Thr Ile 385 390 395 400 Arg Thr Leu Lys Ala Glu Ile Gln Val Gly Leu Gln Pro Gly Gln Gly 405 410 415 Glu Gln Leu Leu Ala Ser Glu Pro Ser Trp Trp Thr Thr Gly Lys Tyr 420 425 430 Gly Ile Phe Gln Tyr Asp Gly Pro Leu 435 440 <210> 12 <211> 260 <212> PRT <213> Artificial Sequence <220> <223> PCSK1N amino acid sequence <400> 12 Met Ala Gly Ser Pro Leu Leu Trp Gly Pro Arg Ala Gly Gly Val Gly 1 5 10 15 Leu Leu Val Leu Leu Leu Leu Gly Leu Phe Arg Pro Pro Ala Leu 20 25 30 Cys Ala Arg Pro Val Lys Glu Pro Arg Gly Leu Ser Ala Ala Ser Pro 35 40 45 Pro Leu Ala Glu Thr Gly Ala Pro Arg Arg Phe Arg Arg Ser Val Pro 50 55 60 Arg Gly Glu Ala Ala Gly Ala Val Gln Glu Leu Ala Arg Ala Leu Ala 65 70 75 80 His Leu Leu Glu Ala Glu Arg Gln Glu Arg Ala Arg Ala Glu Ala Gln 85 90 95 Glu Ala Glu Asp Gln Gln Ala Arg Val Leu Ala Gln Leu Leu Arg Val 100 105 110 Trp Gly Ala Pro Arg Asn Ser Asp Pro Ala Leu Gly Leu Asp Asp Asp 115 120 125 Pro Asp Ala Pro Ala Ala Gln Leu Ala Arg Ala Leu Leu Arg Ala Arg 130 135 140 Leu Asp Pro Ala Ala Leu Ala Ala Gln Leu Val Pro Ala Pro Val Pro 145 150 155 160 Ala Ala Ala Leu Arg Pro Arg Pro Pro Val Tyr Asp Asp Gly Pro Ala 165 170 175 Gly Pro Asp Ala Glu Glu Ala Gly Asp Glu Thr Pro Asp Val Asp Pro 180 185 190 Glu Leu Leu Arg Tyr Leu Leu Gly Arg Ile Leu Ala Gly Ser Ala Asp 195 200 205 Ser Glu Gly Val Ala Ala Pro Arg Arg Leu Arg Arg Ala Ala Asp His 210 215 220 Asp Val Gly Ser Glu Leu Pro Pro Glu Gly Val Leu Gly Ala Leu Leu 225 230 235 240 Arg Val Lys Arg Leu Glu Thr Pro Ala Pro Gln Val Pro Ala Arg Arg 245 250 255 Leu Leu Pro Pro 260 <210> 13 <211> 1555 <212> PRT <213> Artificial Sequence <220> <223> UGGT1 amino acid sequence <400> 13 Met Gly Cys Lys Gly Asp Ala Ser Gly Ala Cys Ala Ala Gly Ala Leu 1 5 10 15 Pro Val Thr Gly Val Cys Tyr Lys Met Gly Val Leu Val Val Leu Thr 20 25 30 Val Leu Trp Leu Phe Ser Ser Val Lys Ala Asp Ser Lys Ala Ile Thr 35 40 45 Thr Ser Leu Thr Thr Lys Trp Phe Ser Thr Pro Leu Leu Leu Glu Ala 50 55 60 Ser Glu Phe Leu Ala Glu Asp Ser Gln Glu Lys Phe Trp Asn Phe Val 65 70 75 80 Glu Ala Ser Gln Asn Ile Gly Ser Ser Asp His Asp Gly Thr Asp Tyr 85 90 95 Ser Tyr Tyr His Ala Ile Leu Glu Ala Ala Phe Gln Phe Leu Ser Pro 100 105 110 Leu Gln Gln Asn Leu Phe Lys Phe Cys Leu Ser Leu Arg Ser Tyr Ser 115 120 125 Ala Thr Ile Gln Ala Phe Gln Gln Ile Ala Ala Asp Glu Pro Pro Pro 130 135 140 Glu Gly Cys Asn Ser Phe Phe Ser Val His Gly Lys Lys Thr Cys Glu 145 150 155 160 Ser Asp Thr Leu Glu Ala Leu Leu Leu Thr Ala Ser Glu Arg Pro Lys 165 170 175 Pro Leu Leu Phe Lys Gly Asp His Arg Tyr Pro Ser Ser Asn Pro Glu 180 185 190 Ser Pro Val Val Ile Phe Tyr Ser Glu Ile Gly Ser Glu Glu Phe Ser 195 200 205 Asn Phe His Arg Gln Leu Ile Ser Lys Ser Asn Ala Gly Lys Ile Asn 210 215 220 Tyr Val Phe Arg His Tyr Ile Phe Asn Pro Arg Lys Glu Pro Val Tyr 225 230 235 240 Leu Ser Gly Tyr Gly Val Glu Leu Ala Ile Lys Ser Thr Glu Tyr Lys 245 250 255 Ala Lys Asp Asp Thr Gln Val Lys Gly Thr Glu Val Asn Thr Thr Val 260 265 270 Ile Gly Glu Asn Asp Pro Ile Asp Glu Val Gln Gly Phe Leu Phe Gly 275 280 285 Lys Leu Arg Asp Leu His Pro Asp Leu Glu Gly Gln Leu Lys Glu Leu 290 295 300 Arg Lys His Leu Val Glu Ser Thr Asn Glu Met Ala Pro Leu Lys Val 305 310 315 320 Trp Gln Leu Gln Asp Leu Ser Phe Gln Thr Ala Ala Arg Ile Leu Ala 325 330 335 Ser Pro Val Glu Leu Ala Leu Val Val Met Lys Asp Leu Ser Gln Asn 340 345 350 Phe Pro Thr Lys Ala Arg Ala Ile Thr Lys Thr Ala Val Ser Ser Glu 355 360 365 Leu Arg Thr Glu Val Glu Glu Asn Gln Lys Tyr Phe Lys Gly Thr Leu 370 375 380 Gly Leu Gln Pro Gly Asp Ser Ala Leu Phe Ile Asn Gly Leu His Met 385 390 395 400 Asp Leu Asp Thr Gln Asp Ile Phe Ser Leu Phe Asp Val Leu Arg Asn 405 410 415 Glu Ala Arg Val Met Glu Gly Leu His Arg Leu Gly Ile Glu Gly Leu 420 425 430 Ser Leu His Asn Val Leu Lys Leu Asn Ile Gln Pro Ser Glu Ala Asp 435 440 445 Tyr Ala Val Asp Ile Arg Ser Pro Ala Ile Ser Trp Val Asn Asn Leu 450 455 460 Glu Val Asp Ser Arg Tyr Asn Ser Trp Pro Ser Ser Leu Gln Glu Leu 465 470 475 480 Leu Arg Pro Thr Phe Pro Gly Val Ile Arg Gln Ile Arg Lys Asn Leu 485 490 495 His Asn Met Val Phe Ile Val Asp Pro Ala His Glu Thr Thr Ala Glu 500 505 510 Leu Met Asn Thr Ala Glu Met Phe Leu Ser Asn His Ile Pro Leu Arg 515 520 525 Ile Gly Phe Ile Phe Val Val Asn Asp Ser Glu Asp Val Asp Gly Met 530 535 540 Gln Asp Ala Gly Val Ala Val Leu Arg Ala Tyr Asn Tyr Val Ala Gln 545 550 555 560 Glu Val Asp Asp Tyr His Ala Phe Gln Thr Leu Thr His Ile Tyr Asn 565 570 575 Lys Val Arg Thr Gly Glu Lys Val Lys Val Glu His Val Val Ser Val 580 585 590 Leu Glu Lys Lys Tyr Pro Tyr Val Glu Val Asn Ser Ile Leu Gly Ile 595 600 605 Asp Ser Ala Tyr Asp Arg Asn Arg Lys Glu Ala Arg Gly Tyr Tyr Glu 610 615 620 Gln Thr Gly Val Gly Pro Leu Pro Val Val Leu Phe Asn Gly Met Pro 625 630 635 640 Phe Glu Arg Glu Gln Leu Asp Pro Asp Glu Leu Glu Thr Ile Thr Met 645 650 655 His Lys Ile Leu Glu Thr Thr Thr Phe Phe Gln Arg Ala Val Tyr Leu 660 665 670 Gly Glu Leu Pro His Asp Gln Asp Val Val Glu Tyr Ile Met Asn Gln 675 680 685 Pro Asn Val Val Pro Arg Ile Asn Ser Arg Ile Leu Thr Ala Glu Arg 690 695 700 Asp Tyr Leu Asp Leu Thr Ala Ser Asn Asn Phe Phe Val Asp Asp Tyr 705 710 715 720 Ala Arg Phe Thr Ile Leu Asp Ser Gln Gly Lys Thr Ala Ala Val Ala 725 730 735 Asn Ser Met Asn Tyr Leu Thr Lys Lys Gly Met Ser Ser Lys Glu Ile 740 745 750 Tyr Asp Asp Ser Phe Ile Arg Pro Val Thr Phe Trp Ile Val Gly Asp 755 760 765 Phe Asp Ser Pro Ser Gly Arg Gln Leu Leu Tyr Asp Ala Ile Lys His 770 775 780 Gln Lys Ser Ser Asn Asn Val Arg Ile Ser Met Ile Asn Asn Pro Ala 785 790 795 800 Lys Glu Ile Ser Tyr Glu Asn Thr Gln Ile Ser Arg Ala Ile Trp Ala 805 810 815 Ala Leu Gln Thr Gln Thr Ser Asn Ala Ala Lys Asn Phe Ile Thr Lys 820 825 830 Met Ala Lys Glu Gly Ala Ala Glu Ala Leu Ala Ala Gly Ala Asp Ile 835 840 845 Ala Glu Phe Ser Val Gly Gly Met Asp Phe Ser Leu Phe Lys Glu Val 850 855 860 Phe Glu Ser Ser Lys Met Asp Phe Ile Leu Ser His Ala Val Tyr Cys 865 870 875 880 Arg Asp Val Leu Lys Leu Lys Lys Gly Gln Arg Ala Val Ile Ser Asn 885 890 895 Gly Arg Ile Ile Gly Pro Leu Glu Asp Ser Glu Leu Phe Asn Gln Asp 900 905 910 Asp Phe His Leu Leu Glu Asn Ile Ile Leu Lys Thr Ser Gly Gln Lys 915 920 925 Ile Lys Ser His Ile Gln Gln Leu Arg Val Glu Glu Asp Val Ala Ser 930 935 940 Asp Leu Val Met Lys Val Asp Ala Leu Leu Ser Ala Gln Pro Lys Gly 945 950 955 960 Asp Pro Arg Ile Glu Tyr Gln Phe Phe Glu Asp Arg His Ser Ala Ile 965 970 975 Lys Leu Arg Pro Lys Glu Gly Glu Thr Tyr Phe Asp Val Val Ala Val 980 985 990 Val Asp Pro Val Thr Arg Glu Ala Gln Arg Leu Ala Pro Leu Leu Leu 995 1000 1005 Val Leu Ala Gln Leu Ile Asn Met Asn Leu Arg Val Phe Met Asn Cys 1010 1015 1020 Gln Ser Lys Leu Ser Asp Met Pro Leu Lys Ser Phe Tyr Arg Tyr Val 1025 1030 1035 1040 Leu Glu Pro Glu Ile Ser Phe Thr Ser Asp Asn Ser Phe Ala Lys Gly 1045 1050 1055 Pro Ile Ala Lys Phe Leu Asp Met Pro Gln Ser Pro Leu Phe Thr Leu 1060 1065 1070 Asn Leu Asn Thr Pro Glu Ser Trp Met Val Glu Ser Val Arg Thr Pro 1075 1080 1085 Tyr Asp Leu Asp Asn Ile Tyr Leu Glu Glu Val Asp Ser Val Val Ala 1090 1095 1100 Ala Glu Tyr Glu Leu Glu Tyr Leu Leu Leu Glu Gly His Cys Tyr Asp 1105 1110 1115 1120 Ile Thr Thr Gly Gln Pro Pro Arg Gly Leu Gln Phe Thr Leu Gly Thr 1125 1130 1135 Ser Ala Asn Pro Val Ile Val Asp Thr Ile Val Met Ala Asn Leu Gly 1140 1145 1150 Tyr Phe Gln Leu Lys Ala Asn Pro Gly Ala Trp Ile Leu Arg Leu Arg 1155 1160 1165 Lys Gly Arg Ser Glu Asp Ile Tyr Arg Ile Tyr Ser His Asp Gly Thr 1170 1175 1180 Asp Ser Pro Pro Asp Ala Asp Glu Val Val Ile Val Leu Asn Asn Phe 1185 1190 1195 1200 Lys Ser Lys Ile Ile Lys Val Lys Val Gln Lys Lys Ala Asp Met Val 1205 1210 1215 Asn Glu Asp Leu Leu Ser Asp Gly Thr Ser Glu Asn Glu Ser Gly Phe 1220 1225 1230 Trp Asp Ser Phe Lys Trp Gly Phe Thr Gly Gln Lys Thr Glu Glu Val 1235 1240 1245 Lys Gln Asp Lys Asp Asp Ile Ile Asn Ile Phe Ser Val Ala Ser Gly 1250 1255 1260 His Leu Tyr Glu Arg Phe Leu Arg Ile Met Met Leu Ser Val Leu Lys 1265 1270 1275 1280 Asn Thr Lys Thr Pro Val Lys Phe Trp Phe Leu Lys Asn Tyr Leu Ser 1285 1290 1295 Pro Thr Phe Lys Glu Phe Ile Pro Tyr Met Ala Asn Glu Tyr Asn Phe 1300 1305 1310 Gln Tyr Glu Leu Val Gln Tyr Lys Trp Pro Arg Trp Leu His Gln Gln 1315 1320 1325 Thr Glu Lys Gln Arg Ile Ile Trp Gly Tyr Lys Ile Leu Phe Leu Asp 1330 1335 1340 Val Leu Phe Pro Leu Val Val Asp Lys Phe Leu Phe Val Asp Ala Asp 1345 1350 1355 1360 Gln Ile Val Arg Thr Asp Leu Lys Glu Leu Arg Asp Phe Asn Leu Asp 1365 1370 1375 Gly Ala Pro Tyr Gly Tyr Thr Pro Phe Cys Asp Ser Arg Arg Glu Met 1380 1385 1390 Asp Gly Tyr Arg Phe Trp Lys Ser Gly Tyr Trp Ala Ser His Leu Ala 1395 1400 1405 Gly Arg Lys Tyr His Ile Ser Ala Leu Tyr Val Val Asp Leu Lys Lys 1410 1415 1420 Phe Arg Lys Ile Ala Ala Gly Asp Arg Leu Arg Gly Gln Tyr Gln Gly 1425 1430 1435 1440 Leu Ser Gln Asp Pro Asn Ser Leu Ser Asn Leu Asp Gln Asp Leu Pro 1445 1450 1455 Asn Asn Met Ile His Gln Val Pro Ile Lys Ser Leu Pro Gln Glu Trp 1460 1465 1470 Leu Trp Cys Glu Thr Trp Cys Asp Asp Ala Ser Lys Lys Arg Ala Lys 1475 1480 1485 Thr Ile Asp Leu Cys Asn Asn Pro Met Thr Lys Glu Pro Lys Leu Glu 1490 1495 1500 Ala Ala Val Arg Ile Val Pro Glu Trp Gln Asp Tyr Asp Gln Glu Ile 1505 1510 1515 1520 Lys Gln Leu Gln Ile Arg Phe Gln Lys Glu Lys Glu Thr Gly Ala Leu 1525 1530 1535 Tyr Lys Glu Lys Thr Lys Glu Pro Ser Arg Glu Gly Pro Gln Lys Arg 1540 1545 1550 Glu Glu Leu 1555 <210> 14 <211> 338 <212> PRT <213> Artificial Sequence <220> <223> AHSA1 amino acid sequence <400> 14 Met Ala Lys Trp Gly Glu Gly Asp Pro Arg Trp Ile Val Glu Glu Arg 1 5 10 15 Ala Asp Ala Thr Asn Val Asn Asn Trp His Trp Thr Glu Arg Asp Ala 20 25 30 Ser Asn Trp Ser Thr Asp Lys Leu Lys Thr Leu Phe Leu Ala Val Gln 35 40 45 Val Gln Asn Glu Glu Gly Lys Cys Glu Val Thr Glu Val Ser Lys Leu 50 55 60 Asp Gly Glu Ala Ser Ile Asn Asn Arg Lys Gly Lys Leu Ile Phe Phe 65 70 75 80 Tyr Glu Trp Ser Val Lys Leu Asn Trp Thr Gly Thr Ser Lys Ser Gly 85 90 95 Val Gln Tyr Lys Gly His Val Glu Ile Pro Asn Leu Ser Asp Glu Asn 100 105 110 Ser Val Asp Glu Val Glu Ile Ser Val Ser Leu Ala Lys Asp Glu Pro 115 120 125 Asp Thr Asn Leu Val Ala Leu Met Lys Glu Glu Gly Val Lys Leu Leu 130 135 140 Arg Glu Ala Met Gly Ile Tyr Ile Ser Thr Leu Lys Thr Glu Phe Thr 145 150 155 160 Gln Gly Met Ile Leu Pro Thr Met Asn Gly Glu Ser Val Asp Pro Val 165 170 175 Gly Gln Pro Ala Leu Lys Thr Glu Glu Arg Lys Ala Lys Pro Ala Pro 180 185 190 Ser Lys Thr Gln Ala Arg Pro Val Gly Val Lys Ile Pro Thr Cys Lys 195 200 205 Ile Thr Leu Lys Glu Thr Phe Leu Thr Ser Pro Glu Glu Leu Tyr Arg 210 215 220 Val Phe Thr Thr Gln Glu Leu Val Gln Ala Phe Thr His Ala Pro Ala 225 230 235 240 Thr Leu Glu Ala Asp Arg Gly Gly Lys Phe His Met Val Asp Gly Asn 245 250 255 Val Ser Gly Glu Phe Thr Asp Leu Val Pro Glu Lys His Ile Val Met 260 265 270 Lys Trp Arg Phe Lys Ser Trp Pro Glu Gly His Phe Ala Thr Ile Thr 275 280 285 Leu Thr Phe Ile Asp Lys Asn Gly Glu Thr Glu Leu Cys Met Glu Gly 290 295 300 Arg Gly Ile Pro Ala Pro Glu Glu Glu Arg Thr Arg Gln Gly Trp Gln 305 310 315 320 Arg Tyr Tyr Phe Glu Gly Ile Lys Gln Thr Phe Gly Tyr Gly Ala Arg 325 330 335 Leu Phe <210> 15 <211> 61 <212> PRT <213> Artificial Sequence <220> <223> MT2A amino acid sequence <400> 15 Met Asp Pro Asn Cys Ser Cys Ala Ala Gly Asp Ser Cys Thr Cys Ala 1 5 10 15 Gly Ser Cys Lys Cys Lys Glu Cys Lys Cys Thr Ser Cys Lys Lys Ser 20 25 30 Cys Cys Ser Cys Cys Pro Val Gly Cys Ala Lys Cys Ala Gln Gly Cys 35 40 45 Ile Cys Lys Gly Ala Ser Asp Lys Cys Ser Cys Cys Ala 50 55 60 <210> 16 <211> 234 <212> PRT <213> Artificial Sequence <220> <223> MESDC2 amino acid sequence <400> 16 Met Ala Ala Ser Arg Trp Ala Arg Lys Ala Val Val Leu Leu Cys Ala 1 5 10 15 Ser Asp Leu Leu Leu Leu Leu Leu Leu Leu Pro Pro Pro Gly Ser Cys 20 25 30 Ala Ala Glu Gly Ser Pro Gly Thr Pro Asp Glu Ser Thr Pro Pro Pro 35 40 45 Arg Lys Lys Lys Lys Asp Ile Arg Asp Tyr Asn Asp Ala Asp Met Ala 50 55 60 Arg Leu Leu Glu Gln Trp Glu Lys Asp Asp Asp Ile Glu Glu Gly Asp 65 70 75 80 Leu Pro Glu His Lys Arg Pro Ser Ala Pro Val Asp Phe Ser Lys Ile 85 90 95 Asp Pro Ser Lys Pro Glu Ser Ile Leu Lys Met Thr Lys Lys Gly Lys 100 105 110 Thr Leu Met Met Phe Val Thr Val Ser Gly Ser Pro Thr Glu Lys Glu 115 120 125 Thr Glu Glu Ile Thr Ser Leu Trp Gln Gly Ser Leu Phe Asn Ala Asn 130 135 140 Tyr Asp Val Gln Arg Phe Ile Val Gly Ser Asp Arg Ala Ile Phe Met 145 150 155 160 Leu Arg Asp Gly Ser Tyr Ala Trp Glu Ile Lys Asp Phe Leu Val Gly 165 170 175 Gln Asp Arg Cys Ala Asp Val Thr Leu Glu Gly Gln Val Tyr Pro Gly 180 185 190 Lys Gly Gly Gly Ser Lys Glu Lys Asn Lys Thr Lys Gln Asp Lys Gly 195 200 205 Lys Lys Lys Lys Glu Gly Asp Leu Lys Ser Arg Ser Ser Lys Glu Glu 210 215 220 Asn Arg Ala Gly Asn Lys Arg Glu Asp Leu 225 230 <210> 17 <211> 529 <212> PRT <213> Artificial Sequence <220> <223> HEXA amino acid sequence <400> 17 Met Thr Ser Ser Arg Leu Trp Phe Ser Leu Leu Leu Ala Ala Ala Phe 1 5 10 15 Ala Gly Arg Ala Thr Ala Leu Trp Pro Trp Pro Gln Asn Phe Gln Thr 20 25 30 Ser Asp Gln Arg Tyr Val Leu Tyr Pro Asn Asn Phe Gln Phe Gln Tyr 35 40 45 Asp Val Ser Ser Ala Ala Gln Pro Gly Cys Ser Val Leu Asp Glu Ala 50 55 60 Phe Gln Arg Tyr Arg Asp Leu Leu Phe Gly Ser Gly Ser Trp Pro Arg 65 70 75 80 Pro Tyr Leu Thr Gly Lys Arg His Thr Leu Glu Lys Asn Val Leu Val 85 90 95 Val Ser Val Val Thr Pro Gly Cys Asn Gln Leu Pro Thr Leu Glu Ser 100 105 110 Val Glu Asn Tyr Thr Leu Thr Ile Asn Asp Asp Gln Cys Leu Leu Leu 115 120 125 Ser Glu Thr Val Trp Gly Ala Leu Arg Gly Leu Glu Thr Phe Ser Gln 130 135 140 Leu Val Trp Lys Ser Ala Glu Gly Thr Phe Phe Ile Asn Lys Thr Glu 145 150 155 160 Ile Glu Asp Phe Pro Arg Phe Pro His Arg Gly Leu Leu Leu Asp Thr 165 170 175 Ser Arg His Tyr Leu Pro Leu Ser Ser Ile Leu Asp Thr Leu Asp Val 180 185 190 Met Ala Tyr Asn Lys Leu Asn Val Phe His Trp His Leu Val Asp Asp 195 200 205 Pro Ser Phe Pro Tyr Glu Ser Phe Thr Phe Pro Glu Leu Met Arg Lys 210 215 220 Gly Ser Tyr Asn Pro Val Thr His Ile Tyr Thr Ala Gln Asp Val Lys 225 230 235 240 Glu Val Ile Glu Tyr Ala Arg Leu Arg Gly Ile Arg Val Leu Ala Glu 245 250 255 Phe Asp Thr Pro Gly His Thr Leu Ser Trp Gly Pro Gly Ile Pro Gly 260 265 270 Leu Leu Thr Pro Cys Tyr Ser Gly Ser Glu Pro Ser Gly Thr Phe Gly 275 280 285 Pro Val Asn Pro Ser Leu Asn Asn Thr Tyr Glu Phe Met Ser Thr Phe 290 295 300 Phe Leu Glu Val Ser Ser Val Phe Pro Asp Phe Tyr Leu His Leu Gly 305 310 315 320 Gly Asp Glu Val Asp Phe Thr Cys Trp Lys Ser Asn Pro Glu Ile Gln 325 330 335 Asp Phe Met Arg Lys Lys Gly Phe Gly Glu Asp Phe Lys Gln Leu Glu 340 345 350 Ser Phe Tyr Ile Gln Thr Leu Leu Asp Ile Val Ser Ser Tyr Gly Lys 355 360 365 Gly Tyr Val Val Trp Gln Glu Val Phe Asp Asn Lys Val Lys Ile Gln 370 375 380 Pro Asp Thr Ile Ile Gln Val Trp Arg Glu Asp Ile Pro Val Asn Tyr 385 390 395 400 Met Lys Glu Leu Glu Leu Val Thr Lys Ala Gly Phe Arg Ala Leu Leu 405 410 415 Ser Ala Pro Trp Tyr Leu Asn Arg Ile Ser Tyr Gly Pro Asp Trp Lys 420 425 430 Asp Phe Tyr Ile Val Glu Pro Leu Ala Phe Glu Gly Thr Pro Glu Gln 435 440 445 Lys Ala Leu Val Ile Gly Gly Glu Ala Cys Met Trp Gly Glu Tyr Val 450 455 460 Asp Asn Thr Asn Leu Val Pro Arg Leu Trp Pro Arg Ala Gly Ala Val 465 470 475 480 Ala Glu Arg Leu Trp Ser Asn Lys Leu Thr Ser Asp Leu Thr Phe Ala 485 490 495 Tyr Glu Arg Leu Ser His Phe Arg Cys Glu Leu Leu Arg Arg Gly Val 500 505 510 Gln Ala Gln Pro Leu Asn Val Gly Phe Cys Glu Gln Glu Phe Glu Gln 515 520 525 Thr <210> 18 <211> 946 <212> PRT <213> Artificial Sequence <220> <223> ITIH2 amino acid sequence <400> 18 Met Lys Arg Leu Thr Cys Phe Phe Ile Cys Phe Phe Leu Ser Glu Val 1 5 10 15 Ser Gly Phe Glu Ile Pro Ile Asn Gly Leu Ser Glu Phe Val Asp Tyr 20 25 30 Glu Asp Leu Val Glu Leu Ala Pro Gly Lys Phe Gln Leu Val Ala Glu 35 40 45 Asn Arg Arg Tyr Gln Arg Ser Leu Pro Gly Glu Ser Glu Glu Met Met 50 55 60 Glu Glu Val Asp Gln Val Thr Leu Tyr Ser Tyr Lys Val Gln Ser Thr 65 70 75 80 Ile Thr Ser Arg Met Ala Thr Thr Met Ile Gln Ser Lys Val Val Asn 85 90 95 Asn Ser Pro Gln Pro Gln Asn Val Val Phe Asp Val Gln Ile Pro Lys 100 105 110 Gly Ala Phe Ile Ser Asn Phe Ser Met Thr Val Asp Gly Lys Thr Phe 115 120 125 Arg Ser Ser Ile Lys Glu Lys Thr Val Gly Arg Ala Leu Tyr Ala Gln 130 135 140 Ala Arg Ala Lys Gly Lys Thr Ala Gly Leu Val Arg Ser Ser Ala Leu 145 150 155 160 Asp Met Glu Asn Phe Arg Thr Glu Val Asn Val Leu Pro Gly Ala Lys 165 170 175 Val Gln Phe Glu Leu His Tyr Gln Glu Val Lys Trp Arg Lys Leu Gly 180 185 190 Ser Tyr Glu His Arg Ile Tyr Leu Gln Pro Gly Arg Leu Ala Lys His 195 200 205 Leu Glu Val Asp Val Trp Val Ile Glu Pro Gln Gly Leu Arg Phe Leu 210 215 220 His Val Pro Asp Thr Phe Glu Gly His Phe Asp Gly Val Pro Val Ile 225 230 235 240 Ser Lys Gly Gln Gln Lys Ala His Val Ser Phe Lys Pro Thr Val Ala 245 250 255 Gln Gln Arg Ile Cys Pro Asn Cys Arg Glu Thr Ala Val Asp Gly Glu 260 265 270 Leu Val Val Leu Tyr Asp Val Lys Arg Glu Glu Lys Ala Gly Glu Leu 275 280 285 Glu Val Phe Asn Gly Tyr Phe Val His Phe Phe Ala Pro Asp Asn Leu 290 295 300 Asp Pro Ile Pro Lys Asn Ile Leu Phe Val Ile Asp Val Ser Gly Ser 305 310 315 320 Met Trp Gly Val Lys Met Lys Gln Thr Val Glu Ala Met Lys Thr Ile 325 330 335 Leu Asp Asp Leu Arg Ala Glu Asp His Phe Ser Val Ile Asp Phe Asn 340 345 350 Gln Asn Ile Arg Thr Trp Arg Asn Asp Leu Ile Ser Ala Thr Lys Thr 355 360 365 Gln Val Ala Asp Ala Lys Arg Tyr Ile Glu Lys Ile Gln Pro Ser Gly 370 375 380 Gly Thr Asn Ile Asn Glu Ala Leu Leu Arg Ala Ile Phe Ile Leu Asn 385 390 395 400 Glu Ala Asn Asn Leu Gly Leu Leu Asp Pro Asn Ser Val Ser Leu Ile 405 410 415 Ile Leu Val Ser Asp Gly Asp Pro Thr Val Gly Glu Leu Lys Leu Ser 420 425 430 Lys Ile Gln Lys Asn Val Lys Glu Asn Ile Gln Asp Asn Ile Ser Leu 435 440 445 Phe Ser Leu Gly Met Gly Phe Asp Val Asp Tyr Asp Phe Leu Lys Arg 450 455 460 Leu Ser Asn Glu Asn His Gly Ile Ala Gln Arg Ile Tyr Gly Asn Gln 465 470 475 480 Asp Thr Ser Ser Gln Leu Lys Lys Phe Tyr Asn Gln Val Ser Thr Pro 485 490 495 Leu Leu Arg Asn Val Gln Phe Asn Tyr Pro His Thr Ser Val Thr Asp 500 505 510 Val Thr Gln Asn Asn Phe His Asn Tyr Phe Gly Gly Ser Glu Ile Val 515 520 525 Val Ala Gly Lys Phe Asp Pro Ala Lys Leu Asp Gln Ile Glu Ser Val 530 535 540 Ile Thr Ala Thr Ser Ala Asn Thr Gln Leu Val Leu Glu Thr Leu Ala 545 550 555 560 Gln Met Asp Asp Leu Gln Asp Phe Leu Ser Lys Asp Lys His Ala Asp 565 570 575 Pro Asp Phe Thr Arg Lys Leu Trp Ala Tyr Leu Thr Ile Asn Gln Leu 580 585 590 Leu Ala Glu Arg Ser Leu Ala Pro Thr Ala Ala Ala Lys Arg Arg Ile 595 600 605 Thr Arg Ser Ile Leu Gln Met Ser Leu Asp His His Ile Val Thr Pro 610 615 620 Leu Thr Ser Leu Val Ile Glu Asn Glu Ala Gly Asp Glu Arg Met Leu 625 630 635 640 Ala Asp Ala Pro Pro Gln Asp Pro Ser Cys Cys Ser Gly Ala Leu Tyr 645 650 655 Tyr Gly Ser Lys Val Val Pro Asp Ser Thr Pro Ser Trp Ala Asn Pro 660 665 670 Ser Pro Thr Pro Val Ile Ser Met Leu Ala Gln Gly Ser Gln Val Leu 675 680 685 Glu Ser Thr Pro Pro Pro His Val Met Arg Val Glu Asn Asp Pro His 690 695 700 Phe Ile Ile Tyr Leu Pro Lys Ser Gln Lys Asn Ile Cys Phe Asn Ile 705 710 715 720 Asp Ser Glu Pro Gly Lys Ile Leu Asn Leu Val Ser Asp Pro Glu Ser 725 730 735 Gly Ile Val Val Asn Gly Gln Leu Val Gly Ala Lys Lys Pro Asn Asn 740 745 750 Gly Lys Leu Ser Thr Tyr Phe Gly Lys Leu Gly Phe Tyr Phe Gln Ser 755 760 765 Glu Asp Ile Lys Ile Glu Ile Ser Thr Glu Thr Ile Thr Leu Ser His 770 775 780 Gly Ser Ser Thr Phe Ser Leu Ser Trp Ser Asp Thr Ala Gln Val Thr 785 790 795 800 Asn Gln Arg Val Gln Ile Ser Val Lys Lys Glu Lys Val Val Thr Ile 805 810 815 Thr Leu Asp Lys Glu Met Ser Phe Ser Val Leu Leu His Arg Val Trp 820 825 830 Lys Lys His Pro Val Asn Val Asp Phe Leu Gly Ile Tyr Ile Pro Pro 835 840 845 Thr Asn Lys Phe Ser Pro Lys Ala His Gly Leu Ile Gly Gln Phe Met 850 855 860 Gln Glu Pro Lys Ile His Ile Phe Asn Glu Arg Pro Gly Lys Asp Pro 865 870 875 880 Glu Lys Pro Glu Ala Ser Met Glu Val Lys Gly Gln Lys Leu Ile Ile 885 890 895 Thr Arg Gly Leu Gln Lys Asp Tyr Arg Thr Asp Leu Val Phe Gly Thr 900 905 910 Asp Val Thr Cys Trp Phe Val His Asn Ser Gly Lys Gly Phe Ile Asp 915 920 925 Gly His Tyr Lys Asp Tyr Phe Val Pro Gln Leu Tyr Ser Phe Leu Lys 930 935 940 Arg Pro 945

Claims (11)

A biomarker composition for predicting and diagnosing proliferative liver cancer having proliferative activity of hepatocarcinoma cells by oleate fatty acids, comprising all 18 proteins listed in the following table.

According to claim 1,
The protein is a biomarker composition for predicting and diagnosing proliferative liver cancer having proliferative activity of hepatocellular carcinoma cells by oleate fatty acid, wherein the expression level is increased in liver cancer cells upon treatment with oleic fatty acids. delete A composition for diagnosing proliferative liver cancer having proliferative activity of hepatic cancer cells by oleate fatty acid, comprising a substance that measures all 18 protein levels listed in the table below.

According to claim 4,
The substance is a composition for diagnosing proliferative liver cancer having proliferative activity of hepatocarcinoma cells by oleate fatty acid, characterized in that the antibody specifically binds to the 18 proteins listed in the table. delete delete (A) measuring the expression level of all 18 proteins listed in the table of claim 1 from biological samples isolated from liver cancer patients; And
(b) measuring the expression level of the protein from the control sample and comparing it with the measurement result of step (a),
Method for providing information for predicting and diagnosing proliferative liver cancer having proliferative activity of liver cancer cells by oleic fatty acids. The method of claim 8,
The biological sample is a liver tissue, liver cells or liver cell culture medium separated from the subject, information providing method for predicting and diagnosing proliferative liver cancer with proliferative activity of liver cancer cells by oleic fatty acids. The method of claim 8,
When the amount of protein expression in the biological sample isolated from the liver cancer patient is increased compared to the control group, further comprising determining that the proliferation of liver cancer cells is promoted by proliferative liver cancer by oleic fatty acid. , Information providing method for predicting and diagnosing proliferative liver cancer having proliferative activity of liver cancer cells by oleic fatty acids. delete

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