2021 Fiscal Year Final Research Report
Development of a Highly Efficient In-Gel Protein Recovery Method for Structural Proteomics
| Project/Area Number |
19K05526
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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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| Review Section |
Basic Section 34020:Analytical chemistry-related
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| Research Institution | Ehime University |
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Principal Investigator |
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| Project Period (FY) |
2019-04-01 – 2022-03-31
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| Keywords | ポリアクリルアミドゲル電気泳動 / トップダウン質量分析 / タンパク質構造解析 |
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| Outline of Final Research Achievements |
Top-down mass spectrometry, a highly sensitive structural analysis method for proteins, enables the ionization of intact proteins and the comprehensive acquisition of their chemical structural information. To effectively obtain structural information of target protein components in biological samples using top-down mass spectrometry, it is essential to first fractionate complex protein components prior to mass spectrometry analysis. In this study, we developed a method for fractionating protein components with high resolution using SDS-polyacrylamide gel electrophoresis and successfully established a sample pre-fractionation workflow to achieve highly sensitive top-down mass spectrometry. It was also found that sample pretreatment by the developed method was effective for structural analysis of protein complexes by chemical cross-linking mass spectrometry.
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| Free Research Field |
プロテオミクス
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| Academic Significance and Societal Importance of the Research Achievements |
SDS-ポリアクリルアミドゲル電気泳動は、生化学・分子生物学研究において既にその有効性が証明され、これまでに多くのライフサイエンス研究で採用されているが、トップダウン質量分析への本格的な導入は進んでいない。本研究で開発したSDSポリアクリルアミドゲル電気泳動をベースとするタンパク質分画技術は、専用装置を必要とせず，簡易かつ低コストなタンパク質回収が可能であり、トップダウン質量分析によるタンパク質構造解析研究の普及を促進することが期待できる。
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