2014 Fiscal Year Final Research Report
Development of N- and C-terminal sequencing method for proteins with posttranslational modification
| Project/Area Number |
24510296
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Living organism molecular science
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| Research Institution | Nara Women's University |
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Principal Investigator |
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| Project Period (FY) |
2012-04-01 – 2015-03-31
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| Keywords | タンパク質の翻訳後修飾 / プロテオミクス / 末端基の修飾 / ケミカルバイオロジー / 化学修飾 / 同位体交換 / 同位体標識 |
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| Outline of Final Research Achievements |
In order to identify mature proteins in which either N- or C-terminus is blocked, a new method for the modification of terminal free amino and carboxyl groups was developed. This method employs reagents containing the tris(trimethoxyphenyl)phosphonium (TMPP) group. In matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS), the TMPP reagents make it possible to detect the derivatized peptides at a greatly enhanced sensitivity, so that the N- and C-terminal sequencing with MALDI-MS can be performed with almost the same sensitivity. In the present study, a new reaction that converts the N-terminal aspartic acid residue to alanine or pyruvate through decarboxylation and subsequent deamination, respectively, was discovered. The possibility of developing a new method that uses this reaction was also pursued. This method was successfully applied to Egyptian archaeological specimens, which dated to 2,400 BG, thereby collagen derived from cow hide could be identified.
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| Free Research Field |
生命有機化学、プロテオミクス、質量分析学
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