2001 Fiscal Year Final Research Report Summary
Development of high-throughput mass-spec based sequencing and its application to global analysis of protein phosphorylation
| Project/Area Number |
11558082
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Structural biochemistry
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| Research Institution | RIKEN (2000-2001)
Fujita Health University (1999) |
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Principal Investigator |
TANIGUCHI Hisaaki RIKEN, Postranlational Modifications and Dynamic Regulation Research Team, Team Leader, 翻訳後修飾による動的調節機構研究チーム, チームリーダー(研究職) (10257636)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUBARA Mamoru RIKEN, Postranlational Modifications and Dynamic Regulation Research Team, Researcher, 翻訳後修飾による動的調節機構研究チーム, 連携研究員 (90288481)
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| Project Period (FY) |
1999 – 2001
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| Keywords | amino acid sequencing / proteome / mass analysis / mass spectrometry / protein / peptide |
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| Research Abstract |
The aims of the research are :
1) To establish, mass mapping method to identify proteins in the genome database with the information obtained by mass spectrometry of peptides generated from 2D gel spots.
2) TO establish a modification-specific method to detect phosphorylated or acetylated peptides.
3) To apply thus established methods to the global analysis of signaling complexes to elucidate the functions of "unknown" proteins.
For these porpoises, we have established an analysis system, in which gel-spotting and excision, mass measurements and database search are performed automatically. To achieve a high-throughput analysis, the sample-pretreatment robot was purchased by the fund, and modified to make automatic desalting and targeting on the sample plate possible. We have also established an analysis system with which specific detection of phosphopeptides are possible by parent scan in the triple-quad mass spectrometer. The high-throughput identification of more than 30 proteins in a single sample became possible by LC/MS analysis. In this way, we could achieve the high-throughput mass sequencing.
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