2023 Fiscal Year Final Research Report
Elucidation of the molecular mechanism of VSP, a voltage-dependent phosphatase that convert electrical signals into chemical signals
| Project/Area Number |
21H02444
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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 | 一般 |
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| Review Section |
Basic Section 43040:Biophysics-related
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| Research Institution | Osaka University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
米澤 康滋 近畿大学, 先端技術総合研究所, 教授 (40248753)
岡村 康司 大阪大学, 大学院医学系研究科, 教授 (80201987)
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| Project Period (FY) |
2021-04-01 – 2024-03-31
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| Keywords | 膜電位 / タンパク質 / 構造生物学 |
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| Outline of Final Research Achievements |
We have conducted research to elucidate the molecular mechanism of the voltage-sensing phosphatase VSP. Various constructs and crystallization conditions were tried for crystal structure determination of the resting state of VSP. Although the structural analysis was not achieved, microcrystals were obtained, and we are continuing to investigate conditions for obtaining better crystals for atomic structure determination. Cryo-EM single particle analysis could not determine the atomic structure due to large domain fluctuations. A combination of electrophysiology, fluorescence measurements, and structural modeling revealed that two hydrophobic residues play an important role in coupling the VSD and the cytoplasmic catalytic region and that the S4 helix and its connecting linker exhibit as a single helix and are involved in activation.
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| Free Research Field |
構造生物学
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| Academic Significance and Societal Importance of the Research Achievements |
電位センサータンパク質VSPの動作機構の解明を目指した研究を進めた。静止状態の構造解析を目指し様々なコンストラクトや結晶化条件を試みた。構造解析には至らなかったが微結晶を得ることができ、今後の研究につなげることができた。また、電気生理学、蛍光測定、構造モデリングを組み合わせることで、2つの疎水性残基が電位センサードメインと細胞質酵素領域のカップリングに重要な役割を果たしていること、S4ヘリックスとそれに繋がるリンカーが1本のヘリックスとして動き活性化に関与するというVSPの活性化機構の一端を明らかにすることができた。
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