2006 Fiscal Year Final Research Report Summary
Proteomics-based approach identifying autoantibodies in Behcet's disease
| Project/Area Number |
17590502
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | Osaka Medical College |
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Principal Investigator |
TAKEUCHI Toru Osaka Medical College, Faculty of Medicine, Research Associate, 医学部, 助手 (10330078)
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| Co-Investigator(Kenkyū-buntansha) |
SHIMIZU Akira Osaka Medical College, Faculty of Medicine, Emeritus Professor, 医学部, 名誉教授 (00028581)
NAKANISHI Toyofumi Osaka Medical College, Faculty of Medicine, Associate Professor, 医学部, 助教授 (10247843)
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| Project Period (FY) |
2005 – 2006
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| Keywords | Behecet's disease / autoantibody / proteome / mass spectrometry / electrophoresis / immunoprecipitation / immunoblotting |
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| Research Abstract |
We identified autoantigens specific for Behcet's disease by proteomics-based approaches. These approaches were the combination between electrophoresis or immunoprecipitation and mass spectrometry (MS).
To detect autoantigens for Behcet's disease, we performed one or two-dimensional electrophoresis using HEp-2 cells and western blotting. There were several spots specific for Behcet's disease, which identified by comparison with sera of normal healthy controls and disease controls.
To identify these spots of interest, the gel spots were excised and each piece of the gel spots was reduced, alkylated, and digested with trypsin. The extracted peptides were analyzed with an UltraFlex. The peptide mass fingerprint was used for protein identification from the tryptic fragment size using the Mascot Search engine based on databases. We identified two proteins, α-enolase and annexin-I, which were reported as autoantigens for Behcet's disease. We have also performed the combination between immunoprecipitation and MS for detecting autoantigens. Other proteins, which are candidate autoantigens, were detected by western blotting and immunoprecipitation, and are now under identification by MS.
The combination between Immunoblotting or immunoprecipitation and MS is a useful tool for identifying autoantigens for autoimmune diseases such as Behect's disease.
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