2023 Fiscal Year Final Research Report
Positive charge-based electrophoresis for protein complexes
| Project/Area Number |
19K05704
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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 37010:Bio-related chemistry
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| Research Institution | Yasuda Women's University |
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Principal Investigator |
Hirano Makoto 安田女子大学, 薬学部, 講師 (60514172)
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| Project Period (FY) |
2019-04-01 – 2024-03-31
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| Keywords | 電気泳動 / 陽イオン |
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| Outline of Final Research Achievements |
In this study, we developed a novel electrophoretic method to natively separate protein complexes containing positive charge such as Ca2+ and Mg2+ ions. Pararosaniline derivatives were synthesized as charge shift molecules. These derivatives were positively charged in a neutral solution. Some of the derivatives could bind to protein complexes derived from several animal tissues. The protein complexes could be electrophoresed by the proposed method. In addition to this, the method could be applied to an electrophoresis of complex composed of a lectin (glycan-recognizing protein) and a glycoprotein.
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| Free Research Field |
生化学
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| Academic Significance and Societal Importance of the Research Achievements |
タンパク質複合体の分離にはゲル濾過も用いられるが、電気泳動の方が分離能は圧倒的に高い。また、複合体の大きさに応じて適切なゲル濾過担体を選択する必要があるが、電気泳動では濃度勾配ゲルを用意すれば幅広い大きさの複合体に対応できる。電気泳動によるタンパク質複合体の分離にはNative PAGEやBlue Native PAGEなどが適用されている。しかし、いずれもタンパク質複合体の負電荷に基づき分離する。本研究にて開発したRN-PAGEでは正電荷に基づき分離する。本法により、これまで困難であった陽イオンを含むタンパク質複合体の分離が可能となり、これまでの負電荷に基づく分離と相補的な手法となり得る。
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