2013 Fiscal Year Final Research Report
Elucidation of the structural basis of the excitation and contraction coupling usiing phase-contrast electron cryotomography
| Project/Area Number |
23570150
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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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| Research Field |
Structural biochemistry
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| Research Institution | National Institute for Physiological Sciences |
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Principal Investigator |
MURATA Kazuyoshi 生理学研究所, 脳機能計測・支援センター, 准教授 (20311201)
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| Co-Investigator(Kenkyū-buntansha) |
KUNIYASU Akihiko 崇城大学, 薬学部, 教授 (90241348)
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| Project Period (FY) |
2011 – 2013
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| Keywords | 高分解能電子顕微鏡解析 / 膜タンパク質 / 興奮収縮連関 / クライオ電子顕微鏡 / 電子線トモグラフィー |
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| Research Abstract |
We extracted enzymatically-active triad membranes from rabbit's skeletal muscle and examined them using electron cryomicroscopy. A lot of proteoliposomes were successfully observed in the sample. Some of them were identified as sarcoplasmic reticulum (SR). We further labeled the dihydropyridine receptor (DHPR) in T-tubules using the anti-beta subunit antibody conjugated fluorescent dyes and gold colloids to detect the triad junction (TJ). Although we can find gold-labels near SRs, The region of TJ has not been found out yet. We will also reconsider the condition of membrane extraction to preserve TJ abundantly, and examine it again in the future.
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