KR20220014516A - Composition for diagnosing Parkinson’s disease comprising an agent for determining level of p32 - Google Patents

Abstract

The present invention relates to a composition for the diagnosis of Parkinson's disease comprising a formulation for determining the level of p32. According to the present invention, the level of the p32 gene or protein can be determined from biological samples of a patient group suspected of having Parkinson's disease, and thus it can be provided as effective information for the prediction and diagnosis of Parkinson's disease.

Description

Composition for diagnosing Parkinson's disease comprising an agent for determining level of p32

The present invention relates to a composition for diagnosing Parkinson's disease comprising an agent for measuring the level of p32.

Parkinson's Disease (PD) is a disease with main symptoms such as tremor, stiffness, slow movement and gait abnormality, and is a neurotransmitter called dopamine in the substantia nigra and corpus striatum of the brain. It is a chronic disease caused by this deficiency. The dopamine is produced in a region called substantia nigra of the brain, and is a substance secreted to control the function of the basal ganglia, which is a very important region for regulating motor functions. Parkinson's disease is estimated to account for approximately 1% of the population over 60 years of age.

Although the exact cause of Parkinson's disease is not known, the 'multifactorial hypothesis' that genetic factors and environmental factors interact with each other is the most widely accepted. Parkinson's disease is divided into sporadic Parkinson's disease and familial Parkinson's disease, and about 10% of them appear as familial Parkinson's disease. In particular, among the various genetic factors that cause Parkinson's disease, Parkinson's disease patients caused by mutations in the Parkin gene appeared the most.

Parkin protein functions to reduce oxidative stress in cells by removing damaged, oxidized and/or irregularly structured proteins inside the cell. In relation to these functions, it is known to be involved in removing damaged mitochondria and preventing the accumulation of alpha-synuclein. When a mutation of the Parkin gene occurs, it loses its properties as an E3-ligase; Inclusion bodies and/or irregular proteins accumulate inside cells, leading to decreased dopamine secretion and apoptosis of dopaminergic neurons. In a study on Parkinson's disease, dopamine secretion was decreased due to inactivation of dopaminergic neurons whose functions of Parkin and PINK1 (PTEN-induced kinase 1) were revealed, and Parkin was overexpressed in fruit flies that did not express PINK1. It was confirmed that Parkinson's disease-related symptoms induced by PINK1, such as mitochondrial dysfunction and degradation of dopaminergic neurons, were recovered. These findings suggest that Parkinson's protein is closely related to Parkinson's-related diseases. Moreover, the degree of Parkinson's protein modification by nitric oxide was 2-3 times higher in the brains of patients with Parkinson's disease than those without Parkinson's disease.

Accordingly, the present inventors completed the present invention by confirming that the level of the p32 protein in the mitochondria is strongly associated with the parkin protein while studying the association between the p32 protein and Parkinson's disease.

Republic of Korea Patent No. 10-2061924

It is an object of the present invention to provide a composition for diagnosing Parkinson's disease comprising an agent for measuring the level of p32 gene, protein, or peptide constituting the protein.

Another object of the present invention is to provide a kit for diagnosing Parkinson's disease comprising the composition for diagnosing Parkinson's disease.

Then, another object of the present invention is to provide a method for providing information for prediction and judgment of Parkinson's disease.

Finally, another object of the present invention is to provide a method for screening a candidate for a therapeutic agent for Parkinson's disease.

In order to achieve the above object, the present invention provides a composition for diagnosing Parkinson's disease comprising an agent for measuring the level of the p32 gene, protein, or peptide constituting the protein.

In addition, there is provided a kit for diagnosing Parkinson's disease comprising the composition for diagnosing Parkinson's disease.

Subsequently, the first step of measuring the expression level of the p32 gene or protein from the biological sample; And, it provides a method of providing information for prediction and determination of Parkinson's disease, comprising the second step of comparing the expression level of the gene or protein with the expression level of the gene or protein from a normal control sample.

Finally, step 1 of treating a sample isolated from a Parkinson's disease patient with a candidate drug; and, analyzing the p32 gene activity or protein expression level in the sample treated with the candidate drug.

It was confirmed that the degradation of the p32 protein of the present invention is induced by the Parkin protein, which is closely related to Parkinson's disease. Therefore, by measuring the level of the p32 gene or protein from a biological sample of a patient group suspected of having Parkinson's disease through the present invention, it can be provided as effective information for prediction and diagnosis of Parkinson's disease.

1 is a photograph showing the Western blot results of p32 protein according to arginase II (Arginase II) and Parkin protein mutations.
Figure 2 is a photograph showing whether the p32 protein degradation of the parkin protein according to the p32 mutation.
Figure 3 is a graph comparing the amount of reactive oxygen production according to the p32 mutation.

The present inventors confirmed that the degradation of p32 protein is induced by Parkin protein, which is closely related to Parkinson's disease, while researching an accurate early diagnosis method for Parkinson's disease, one of the neurodegenerative diseases, and p32 protein is When not degraded by protein, it was confirmed that oxidative stress and mitochondrial damage suggested as a mechanism of Parkinson's disease occurred, thereby completing the present invention.

Hereinafter, the present invention will be described in more detail.

The term "Parkinson's disease" as used in the present invention is a disease in which nerve degeneration (degeneration) is caused by the gradual loss of dopamine neurons distributed in the substantia nigra of the brain. It is a chronic progressive degenerative disease of the nervous system characterized by (slow movement) and postural instability. The cause of such Parkinson's disease is still unknown, so it is difficult to predict or determine Parkinson's disease.

As used herein, the term "diagnosis" refers to determining the susceptibility of an individual to a specific disease or disorder, determining whether an individual currently has a specific disease or disorder, or providing information on the efficacy of treatment This includes monitoring the condition of the object to For the purpose of the present invention, diagnosis is to determine whether or not Parkinson's disease has occurred or the stage of onset of Parkinson's disease.

The present invention provides a composition for diagnosing Parkinson's disease, comprising an agent for measuring the level of the p32 gene, protein, or peptide constituting the protein.

In addition, the p32 protein may be composed of all or part of a peptide having a function of interacting with the parkin protein, and includes functional equivalents and variants of the p32 protein.

In addition, the peptide constituting the p32 protein refers to a fragment of the p32 protein, and any peptide constituting the p32 protein having a function of interacting with the parkin protein is included without limitation.

In addition, the functional equivalent of the protein includes proteins and polypeptides that do not entirely alter the activity of the protein molecule, and proteins capable of performing the same functionally are included in the scope of the present invention. The variant refers to a protein having a sequence different from the p32 native amino acid sequence by deletion, insertion, non-conservative or conservative substitution of one or more amino acid residues, or a combination thereof. For example, one or more amino acids in the amino acid sequence are deleted, substituted, or added, and 95% or more, preferably 98% or more, more preferably 99% or more identity to the amino acid sequence is referred to. . Herein, "identity" means that when two amino acid sequences are aligned to the highest degree of identity with or without introducing a gap, the other amino acid sequence with respect to the total number of amino acid residues in one amino acid sequence, including the number of gaps. refers to the ratio (%) of the number of identical amino acid residues of In addition, "several" means an integer of 2 to 10, for example, an integer of 2 to 7, 2 to 5, 2 to 4, or 2 to 3. Specific examples of the natural variant include variants based on polymorphisms such as SNP (single nucleotide polymorphism), splice variants, and the like. Preferably, the substitution is a conservative amino acid substitution. This is because, if it is a conservative amino acid substitution, it may have a structure or property substantially equivalent to that of p32 having the amino acid sequence. Conservative amino acids are non-polar amino acids (glycine, alanine, phenylalanine, valine, leucine, isoleucine, methionine, proline, tryptophan) and polar amino acids (amino acids other than non-polar amino acids), charged amino acids (acidic amino acids (aspartic acid, glutamic acid) and basic amino acids). Amino acids (arginine, histidine, lysine)) and uncharged amino acids (amino acids other than charged amino acids), aromatic amino acids (phenylalanine, tryptophan, tyrosine), branched amino acids (leucine, isoleucine, valine) and aliphatic amino acids (glycine, alanine, leucine) , isoleucine, valine) and the like are known.

In addition, it may include a protein whose structural stability to heat, pH, etc. of the protein is increased or protein activity is increased due to mutation or modification in the amino acid sequence.

In addition, the agent for measuring the level of the gene may be a primer or probe that specifically binds to a gene, but is not limited thereto.

In addition, the measurement of the gene level is reverse transcription polymerase chain reaction (RT-PCR), competitive polymerase chain reaction, real-time polymerase chain reaction, nuclease protection assay (RNase, S1 nuclease assay), in situ hybridization, DNA microarray Usage, Northern blot, Western blot, ELISA (Enzyme Linked Immuno Sorbent Assay), radioimmunoassay, immunodiffusion assay, immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, FACS, mass spectrometry and protein It may be performed by one or more methods selected from the group consisting of microarray methods, but is not limited thereto.

In addition, the agent for measuring the level of the protein or peptide constituting the protein may be any one selected from the group consisting of an antibody, antisense RNA, aptamer, and compound specific for the protein or fragment peptide. , but not limited thereto.

As used herein, the term “antibody” refers to a specific protein molecule directed against an antigenic site. For the purposes of the present invention, an antibody refers to an antibody that specifically binds to the p32 protein, and includes both polyclonal antibodies, monoclonal antibodies and recombinant antibodies.

In addition, the antibody includes functional fragments of the antibody molecule as well as complete forms having two full-length light chains and two full-length heavy chains. A functional fragment of an antibody molecule refers to a fragment having at least an antigen-binding function, and includes Fab, F(ab'), F(ab') 2 and Fv.

In addition, the polyclonal antibody can be prepared by injecting a protein expressed by the gene or a fragment thereof as an immunogen into an external host according to a method known to those skilled in the art. External hosts include mammals such as mice, rats, sheep or rabbits. The immunogen is injected by intramuscular, intraperitoneal or subcutaneous injection, and is generally administered together with an adjuvant to increase antigenicity. Serum is routinely collected from an external host to collect sera showing improved titer and antigen specificity, or to separate and purify antibodies therefrom.

In addition, the monoclonal antibody can be prepared by a technique for generating an immortalized cell line by fusion known to those skilled in the art. For example, a mouse is immunized with the protein expressed by the gene, or a peptide is synthesized and combined with bovine serum albumin to immunize the mouse. An antigen-producing B lymphocyte isolated from a mouse is fused with a human or mouse myeloma to generate an immortalized hybridoma, and the hybridoma cell is used in an indirect enzyme-linked immunosorbent assay (enzyme- Using linked immunoabsorbent assays, ELISA) to check whether monoclonal antibodies are generated, select positive clones and culture them, then separate and purify the antibodies or inject them into the abdominal cavity of rats and collect ascites to produce monoclonal antibodies. .

In addition, the protein level is measured by Western blot, ELISA (enzyme linked immunosorbent assay, ELISA), radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immune diffusion method, rocket ( rocket) 1 selected from the group consisting of immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, flow cytometry (Fluorescence Activated Cell Sorter, FACS), and protein chip It may be carried out in more than one way, but is not limited thereto.

Next, the present invention provides a kit for diagnosing Parkinson's disease comprising the composition for diagnosing Parkinson's disease.

In the present invention, the term "kit" refers to a primer, probe, or antibody, antisense RNA, aptamer or compound that selectively recognizes p32 protein for measuring the expression level of p32 gene, as well as one or more other component compositions suitable for analysis methods , solutions, or devices.

In addition, the present invention comprises the steps of measuring the expression level of a p32 gene or protein from a biological sample; And, it provides a method of providing information for prediction and determination of Parkinson's disease, comprising the step of comparing the expression level of the gene or protein with the expression level of the gene or protein from a normal control sample.

In addition, the biological sample may be selected from the group consisting of tissue, whole blood, serum, and plasma isolated from a Parkinson's disease patient, and may include saliva, sputum, cerebrospinal fluid, or urine.

In addition, the method may further include predicting and judging Parkinson's disease when the expression level of the p32 gene or protein in the biological sample is higher than that of the normal control.

In addition, the normal control refers to a biological sample isolated from an individual not diagnosed with Parkinson's disease, and the protein antibody and antisense RNA from a sample isolated from a normal individual and a sample isolated from an individual for which a diagnosis of Parkinson's disease is to be predicted. Alternatively, by comparing the aptamer level, it is possible to accurately predict whether an individual suspected of having Parkinson's disease develops Parkinson's disease.

When the expression level of the p32 gene or protein of the biological sample is significantly higher than the expression level of the p32 gene or protein of the normal control, the onset or high risk of Parkinson's disease may be predicted.

In addition, when the relative intensity of the biological sample for the p32 gene or protein in the mitochondria of the normal control is 1.5 to 2.5, the method is predicting or judging that Parkinson's disease has occurred or the risk of developing Parkinson's disease is high. .

The relative intensity indicates the relative expression level of the p32 gene or protein.

Finally, the present invention comprises the steps of treating a candidate drug to a sample isolated from a Parkinson's disease patient; and analyzing the level of p32 gene activity or protein expression in the sample treated with the candidate drug.

In addition, the candidate drug includes, without limitation, a substance expected to be effective in the prevention or treatment of Parkinson's disease as a newly synthesized or known compound.

In addition, the activity of the gene is reverse transcription polymerase chain reaction (RT-PCR), competitive polymerase chain reaction, real-time polymerase chain reaction, nuclease protection assay (RNase, S1 nuclease assay), in situ hybridization, DNA microarray method , Northern blot, Western blot, ELISA (Enzyme Linked Immuno Sorbent Assay), radioimmunoassay, immunodiffusion, immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, FACS, mass spectrometry and protein micro It may be performed by one or more methods selected from the group consisting of array methods, but is not limited thereto.

In addition, the expression of the protein is Western blot, ELISA (enzyme linked immunosorbent assay, ELISA), radioimmunoassay (RIA), radioimmunodiffusion method (radioimmunodiffusion), Ouchterlony immune diffusion method, rocket (rocket) ) One selected from the group consisting of immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, flow cytometry (Fluorescence Activated Cell Sorter, FACS), and protein chip It may be performed by the above method, but is not limited thereto.

In addition, Parkinson's disease symptoms (e.g., decreased dopaminergic nerve, cranial nerve inflammation, increased LPS level in each tissue, accumulation (or increase) of alpha-synuclein, or exercise If alleviation of loss of ability (or impairment) is observed, it is determined that the drug is effective for the prevention or treatment of Parkinson's disease.

Hereinafter, embodiments of the present invention will be described in more detail with reference to the accompanying drawings. However, the following examples are only intended to embody the contents of the present invention, and the present invention will not be limited thereby.

<Example 1> Materials and methods

1-1. cell culture

Human umbilical vein endothelial cells (HUVEC) were collected by tying the umbilical cord with a plastic two-way valve and injecting Type II collagenase (250 units/ml in PBS). This was suspended in a growth medium (20% FBS, 1X penicillin/streptomycin, 5 units/ml heparin, 6 ng/ml bFGF in M199 medium), aliquoted into a 100 mm culture dish, and cultured at 37°C, 5% CO2 environment.

1-2. laboratory animal

This experiment was performed in accordance with the guidelines for the care and use of laboratory animals under the approval of the Institutional Review Board of Kangwon National University.

Mice in which the p32 gene was specifically targeted for vascular endothelial cells were used. The genetic background is C57BL/6J, and it was produced using the Tie promoter, and when recombination occurs by the Cre gene, a recombinant protein, the p32 gene is hit.

A mouse having the p32 flox/flox gene and a mouse having the tie Cre gene were crossed to obtain a p32 flox/-Cre+ mouse, and the p32 flox/flox Cre+ mouse was produced by crossbreeding again. It was confirmed that the prepared mice expressed endothelial cell-specific Cre recombinase and had a deletion in the p32 gene.

1-3. Western blot analysis

Cell extraction and tissue extraction were collected using SDS-PAGE, and band densitometry was analyzed using NIH Image J.

1-4. Measurement of reactive oxygen species (ROS)

After the mouse aorta was isolated, it was cultured in Dulbecco's modified Eagle's medium containing 2% fetal bovine serum (FBS) and 1X P/S (penicillin/streptomycin) at 37°C and 5% CO2 conditions for 18 hours. Set the temperature of the heating block of the fluorescence microscope to 34.5°C, preheat it, and then preheat HEPES buffer (NaCl 7.92 g, KCl 0.44 g, MgCl2 0.12 g, HEPES 2.77 g, glucose 2.07 g (pH7.4), CaCl2 0.25 g). , distilled water 1 L) was spread and fixed in a dish filled with blood vessels. Then, for reactive oxygen (ROS) measurement, dihydroethidium (DHE, 5 uM) was treated and then intensified camera (Luca 658M-TL) at DHE (470/580 nm) wavelength and a custom image acquisition program (Cell software, Olympus) was used to determine the fluorescence value. When measuring reactive oxygen species (ROS), MnTBAP (10-5 mol/L) was treated to measure the fluorescence value and used as a control, and the amount of change in fluorescence value was measured through linear regression analysis (version7.5, OriginLab Corp, Northampton, MA). It was decided.

1-5. Mitochondrial fractionation (fractionation)

Cells were washed in subcellular fraction buffer solution [250 mM sucrose, 20 mM HEPES, pH 7.4, 10 mM KCl, 1.5 mM MgCl2, 1 mM EDTA, 1 mM EGTA, protease inhibitors (Roche Co.)] for 3 min. The cells were disrupted twice, and centrifuged at 1000 g for 10 minutes to remove cell remnants and undisrupted cells. The supernatant was centrifuged at 21000 g at 4° C. for 45 minutes, and cytoplasmic and mitochondrial fractions containing 20 μg protein were used for Western blot.

<Example 2> Degradation of p32 protein by Parkin protein

2-1. p32 proteolysis of parkin protein

To confirm that the p32 protein is degraded by the parkin protein, the p32 protein level was measured after mutating the arginase II and parkin proteins in the mitochondria and the cytosol. The amino acid sequence of the p32 protein is shown in Table 1.

siRNA for genes for arginase Ⅱ (Arginase Ⅱ), PINK1 (PTEN-induced kinase 1) and parkin (Parkin) protein were prepared and directly treated with HUVEC. Arginase II antibody, PINK1 antibody (Cat. No. #PA5-13402, invitrogen), Parkin antibody (Cat. No. ab15494, abcam), p32 antibody (Cat. No. ab101267, abcam), HSP60 antibody and actin ( Actin) antibody was used to perform Western blot analysis. The siRNA used in the present invention is shown in Table 2 below. The relative intensity for the p32 protein was calculated using Equation 1 below, and the results are shown in FIG. 1 .

1 is a photograph showing the Western blot results of p32 protein according to arginase II (Arginase II) and Parkin protein mutations.

As shown in Figure 1, arginase II (Arginase II) siRNA induced p32 protein in both mitochondria and cytosol. However, when mutated using PINK1 (PTEN-induced kinase 1) siRNA and Parkin siRNA, it was confirmed that the p32 protein level, which was reduced by Arginase II siRNA, was increased again. In addition, PINK1 (PTEN-induced kinase 1) and Parkin (Parkin) are proteins closely related to mitochondrial quality control by identifying and destroying damaged mitochondria, and mutations in their genes or proteins are known to cause Parkinson's disease. When the amount of PINK 1 or Parkin protein induced by Parkinson's disease was decreased with siRNA, the relative intensity of the p32 protein in mitochondria in the experimental group was 1.9, 2.1, and 1.9, respectively, compared to the normal control group. It was confirmed that the level significantly increased. These results mean that the p32 protein was degraded by the parkin protein, and the level of the p32 gene or protein closely related to the parkin protein can be measured to be usefully used to predict or determine Parkinson's disease.

Protein Sequence p32 MLHTDGDKAF VDFLSDEIKE ERKIQKHKTL PKMSGGWELE LNGTEAKLVR KVAGEKITVT FNINNSIPPT FDGEEEPSQG QKVEEQEPEL TSTPNFVVEV IKNDDGKKAL VLDCHYPEDE WALYDHITEMEAESD IFSIREVSFQ STGESEWKD HQ FLYELDCHYPEDE WALYDHITEDEAESD IFSIREVSFQ STGESEWKK ADE K STGESEWK

name target sequence sip32 p32 TGT CTC CGT CGG TGT GCA GC siparkin parkin UUU ACA GAG AAA CAC CUU GUC AAU CCG ACU CUC UCC AUC AGA AGG GUU siPINK 1 PINK 1 AAG GAU GUU GUC GGA UUU UUC AUA GGC GAU GGC UCC

2-2. p32 proteolysis of Parkin protein according to p32 mutation

In order to check whether the degradation of p32 protein is induced by the parkin protein, p32 was mutated and the binding between p32 and ubiquitin according to the presence or absence of Parkin protein was investigated.

A plasmid was prepared in which lysine residues 154 and 220 of the p32 protein sequence were substituted with arginine, and the normal p32 plasmid and the mutant p32 plasmid were treated with HUVEC. The mutated p32 protein has a Flag tag. Immunoprecipitation was performed on the p32 protein using a p32 antibody or an antibody against Flag, and the protein was isolated by SDS-PAGE, followed by Western blot analysis using an antibody against ubiquitin. Whether ubiquitination was induced by the Parkin protein was compared with the control group through Western blot analysis. The results are shown in FIG. 2 .

Figure 2 is a photograph showing whether the p32 protein degradation of the parkin protein according to the p32 mutation.

As shown in FIG. 2 , it was confirmed that the normal p32 protein was decomposed by binding to ubiquitin by the parkin protein. However, it was confirmed that when the mutation was induced with respect to the p32 protein, the p32 protein was not ubiquitinated by the parkin protein and thus not degraded compared to the case where the mutation was not induced. These results mean that the p32 protein is degraded by the Parkinson protein, and can be usefully used to predict or determine Parkinson's disease by measuring the level of the p32 gene or protein.

<Example 3> Effect of p32 protein on active oxygen production amount

To determine the effect of p32 protein on the amount of reactive oxygen species produced, the amount of reactive oxygen production was measured by hitting p32 in mice.

Endothelial cells-specifically, aortic cells from mice lacking p32 were isolated, and immunohistochemistry was performed using DHE, a fluorescent dye for reactive oxygen species, followed by a fluorescence microscope (Olympus) connected with Clara digital camera (Andor Technol). ) was used to analyze active oxygen production. MnTBAP was used as a positive control as a reactive oxygen scavenger.

The results are shown in FIG. 3 . Figure 3 is a graph comparing the amount of reactive oxygen production according to the p32 mutation.

As shown in FIG. 3 , as a result of measuring the amount of reactive oxygen species produced in p32 knockout mice, it was confirmed that the amount of reactive oxygen production was reduced by about 50% compared to wild-type mice in which p32 was not mutated. Oxidative stress is one of several mechanisms of Parkinson's disease. It means that it can be usefully used to predict or judge a disease.

As described above, although specific embodiments of the present invention have been described in detail, those skilled in the art who understand the spirit of the present invention may add, change, delete, etc. other components within the scope of the same spirit, and other degenerate inventions However, other embodiments included within the scope of the present invention may be easily proposed. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention is indicated by the claims described later rather than the above detailed description, and all changes or modifications derived from the meaning and scope of the claims and their equivalents are included in the scope of the present invention. should be interpreted as

Claims (12)

A composition for diagnosing Parkinson's disease, comprising an agent for measuring the level of the p32 gene, protein, or peptide constituting the protein.
The composition for diagnosing Parkinson's disease according to claim 1, wherein the agent for measuring the gene level comprises a primer or probe that specifically binds to a gene.
The composition for diagnosing Parkinson's disease according to claim 1, wherein the agent for measuring the level of the protein or peptide constituting the protein comprises an antibody, antisense RNA or aptamer specific for the protein or peptide.
According to claim 1, wherein the measurement of the gene level is reverse transcription polymerase chain reaction (RT-PCR), competitive polymerase chain reaction, real-time polymerase chain reaction, nuclease protection assay (RNase, S1 nuclease assay), in situ hybridization method , DNA microarray method, Northern blot, Western blot, ELISA (Enzyme Linked Immuno Sorbent Assay), radioimmunoassay, immunodiffusion method, immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, FACS, mass spectrometry (Mass spectrometry) and a protein microarray method, which is performed by at least one method selected from the group consisting of, a composition for diagnosing Parkinson's disease.
According to claim 1, wherein the measurement of the level of the protein or peptides constituting the protein is Western blot, ELISA (enzyme linked immunosorbent assay, ELISA), radioimmunoassay (RIA), radioimmunodiffusion method (radioimmunodiffusion), OAK Ouchterlony immunodiffusion method, rocket immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay (Immunoprecipitation Assay), complement fixation assay (Complement Fixation Assay), flow cytometry (Fluorescence Activated Cell Sorter, FACS), protein chip (protein chip) that is performed by one or more methods selected from the group consisting of, Parkinson's disease diagnostic composition.
A kit for diagnosing Parkinson's disease comprising the composition for diagnosing Parkinson's disease of claim 1.
measuring the expression level of the p32 gene or protein from the biological sample; and,
A method of providing information for predicting and judging Parkinson's disease, comprising comparing the expression level of the gene or protein with the expression level of the gene or protein from a normal control sample.
The method of claim 7, wherein the biological sample is selected from the group consisting of tissue, whole blood, serum and plasma isolated from a Parkinson's disease patient.
According to claim 7, wherein the method further comprises the step of predicting and judging Parkinson's disease when the expression level of the p32 gene or protein in the biological sample is higher than that of the normal control sample. Information for predicting and judging Parkinson's disease How to provide.
treating a sample isolated from a Parkinson's disease patient with a candidate drug; and,
A screening method for a candidate for Parkinson's disease, comprising analyzing the level of p32 gene activity or protein expression in the sample treated with the candidate drug.
11. The method of claim 10, wherein the activity of the gene is reverse transcription polymerase chain reaction (RT-PCR), competitive polymerase chain reaction, real-time polymerase chain reaction, nuclease protection assay (RNase, S1 nuclease assay), in situ hybridization, DNA microarray method, Northern blot, Western blot, ELISA (Enzyme Linked Immuno Sorbent Assay), radioimmunoassay, immunodiffusion method, immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, FACS, mass spectrometry ) and a method of screening a candidate for Parkinson's disease treatment, which is performed by at least one method selected from the group consisting of a protein microarray method.
11. The method of claim 10, wherein the expression of the protein is Western blot, ELISA (enzyme linked immunosorbent assay, ELISA), radioimmunoassay (RIA), radioimmunodiffusion (radioimmunodiffusion), Ouchterlony (Ouchterlony) immune diffusion method , Rocket immunoelectrophoresis, tissue immunostaining, Immunoprecipitation Assay, Complement Fixation Assay, Flow Cytometry (Fluorescence Activated Cell Sorter, FACS), a group consisting of a protein chip A screening method for Parkinson's disease drug candidates, which is performed by one or more methods selected from

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