論文 - 詳細
| RRC ID | 66075 |
|---|---|
| 著者 | Katoh TA, Daiho T, Yamasaki K, Danko S, Fujimura S, Suzuki H. |
| タイトル | Angle change of the A-domain in a single SERCA1a molecule detected by defocused orientation imaging. |
| ジャーナル | Sci Rep |
| Abstract |
The sarcoendoplasmic reticulum Ca2+-ATPase (SERCA) transports Ca2+ ions across the membrane coupled with ATP hydrolysis. Crystal structures of ligand-stabilized molecules indicate that the movement of actuator (A) domain plays a crucial role in Ca2+ translocation. However, the actual structural movements during the transitions between intermediates remain uncertain, in particular, the structure of E2PCa2 has not been solved. Here, the angle of the A-domain was measured by defocused orientation imaging using isotropic total internal reflection fluorescence microscopy. A single SERCA1a molecule, labeled with fluorophore ReAsH on the A-domain in fixed orientation, was embedded in a nanodisc, and stabilized on Ni-NTA glass. Activation with ATP and Ca2+ caused angle changes of the fluorophore and therefore the A-domain, motions lost by inhibitor, thapsigargin. Our high-speed set-up captured the motion during EP isomerization, and suggests that the A-domain rapidly rotates back and forth from an E1PCa2 position to a position close to the E2P state. This is the first report of the detection in the movement of the A-domain as an angle change. Our method provides a powerful tool to investigate the conformational change of a membrane protein in real-time. |
| 巻・号 | 11(1) |
| ページ | 13672 |
| 公開日 | 2021-7-1 |
| DOI | 10.1038/s41598-021-92986-3 |
| PII | 10.1038/s41598-021-92986-3 |
| PMID | 34211016 |
| PMC | PMC8249593 |
| MeSH | Adenosine Triphosphate / metabolism Animals COS Cells Calcium / metabolism Chlorocebus aethiops Models, Molecular Protein Domains Rabbits Sarcoplasmic Reticulum Calcium-Transporting ATPases / chemistry* Sarcoplasmic Reticulum Calcium-Transporting ATPases / metabolism |
| IF | 3.998 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 4 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | COS-1(RCB0143) |