High resolution and dynamic structural analysis of the entire mammalian FoF1ATP synthase
| Project/Area Number |
19K15749
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 38030:Applied biochemistry-related
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| Research Institution | Kyoto University |
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Principal Investigator |
Jiko Chimari 京都大学, 複合原子力科学研究所, 特別研究員(RPD) (70585976)
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| Project Period (FY) |
2019-04-01 – 2023-03-31
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| Project Status |
Completed (Fiscal Year 2022)
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| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2022: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2021: ¥520,000 (Direct Cost: ¥400,000、Indirect Cost: ¥120,000)
Fiscal Year 2020: ¥520,000 (Direct Cost: ¥400,000、Indirect Cost: ¥120,000)
Fiscal Year 2019: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
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| Keywords | Fo F1ATP合成酵素 / ミトコンドリア / 単粒子構造解析 / ATP合成酵素 / 膜タンパク質複合体 / 膜蛋白質複合体 / 精製 / 界面活性剤 / 膜タンパク質 |
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| Outline of Research at the Start |
哺乳動物のミトコンドリアFoF1ATP合成酵素の全体構造を決定するため、本研究では、安定で無傷な状態で本酵素を精製するため、独自に開発してきたカラムフリー精製について精製純度をさらに高め、全体構造を解析する。構造解析には①三次元結晶に対するＸ線回折と②単粒子に対するクライオ電顕により、高分解能で構造を決定する、及び③自由に回転する状態にあるものについてクライオ電顕により立体構造変化を捉える、という三つの手段を用い、本酵素のエネルギー伝達メカニズムの解明を目指す。
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| Outline of Final Research Achievements |
Monomers, tetramers, and oligomers of mammalian mitochondrial FoF1ATP synthase, which are extremely unstable due to their flexibility with rotation, were stably purified without dissociation of subunits per form, and highly monodisperse cryogrids were successfully prepared. Single-particle structural analysis using cryo-electron microscopy led to a structure of the entire I F1-bound tetramer at approximately 7angstrom resolution.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究は、ミトコンドリアの内膜に存在し、哺乳動物の生命活動を維持するために必要なATPエネルギーの95％以上を生産し、また近年の研究によってミトコンドリア膜透過性遷移孔（PTP）として機能することが明らかになってきているFoF1ATP 合成酵素の高分解能構造解析である。その詳細な構造は、本酵素が関与する疾患の創薬研究の基盤となる。
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Report
(5 results)
Research Products
(5 results)
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[Journal Article] Inhibition of ATP-synthase reverse-activity restores energy homeostasis in mitochondrial pathologies.2023
Author(s)
Perez, R., Beninca, C., Fernandez del Rio, L., Baghdasarian, C. Shu S., Zanette, V., Gerle, C., Jiko, C., Khairallah, R., Khan, S., Pacheco, D. R. F., Shabane, B., Erion, K., Masand, R.,S. Dugar, S., Ghenoiu, C., Schreiner, G., Stiles, L., Liesa, M., Shirihai, O.S.
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Journal Title
EMBO Journal
Volume: e111699
Issue: 10
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] Rapid and Highly Stable Membrane Reconstitution by LAiR Enables the Study of Physiological Integral Membrane Protein Functions.2023
Author(s)
Hernandez, A., Asseri, A.H., Purugganan, A.J., Jiko, C., deRam, C., Lill, H., Pabst, M., Mitsuoka, K., Gerle, C., Bald, D., McMillan,D.G.G.
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Journal Title
ACS Central Science
Volume: 9
Issue: 3
Pages: 494-507
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] Purified F-ATP synthase forms a Ca2+dependent high-conductance channel matching the mitochondrial permeability transition pore.2019
Author(s)
Urbani A,Giorgio V,Carrer A, Franchin C,Arrigoni G, Jiko C,Abe K,Maeda S, Sginzawa-Itoh K, Bogers JF, McMillan DG, Gerle C, Szabo I, Bernardi P
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Journal Title
Nature communications
Volume: 4341
Issue: 1
Pages: 4341-4341
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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