Studies by molecular biological methods on proton pumping mechanisms of bovine heart and bacterial cytochrome c oxidases
Project/Area Number |
21370073
|
Research Category |
Grant-in-Aid for Scientific Research (B)
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Biophysics
|
Research Institution | University of Hyogo |
Principal Investigator |
SHIMADA Hideo 兵庫県立大学, 大学院・生命理学研究科, 教授 (80095611)
|
Co-Investigator(Renkei-kenkyūsha) |
KATAYAMA Yukie 兵庫県立大学, 大学院・生命理学研究科, 特任講師 (70464998)
AMINAKA Ryohta 兵庫県立大学, 大学院・生命理学研究科, 特任助教
|
Project Period (FY) |
2009 – 2011
|
Project Status |
Completed (Fiscal Year 2011)
|
Budget Amount *help |
¥18,980,000 (Direct Cost: ¥14,600,000、Indirect Cost: ¥4,380,000)
Fiscal Year 2011: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2010: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2009: ¥8,320,000 (Direct Cost: ¥6,400,000、Indirect Cost: ¥1,920,000)
|
Keywords | タンパク質 / 核酸の構造 / 動態 / 機能 / チトクロムc酸化酵素 / プロトンポンプ / 部位特異的アミノ酸置換 / ウシ心筋 / 無細胞タンパク質合成系 / Paracoccus denitrificans / N-acetylcysteine / HeLa細胞 / 膜タンパク質複合体 |
Research Abstract |
Function of a putative proton transfer pathway, D-pathway of bovine heart cytochrome c oxidase (CcO) was analyzed by site-directed mutagenesis employing HeLa cell expression system we have developed to produce bovine/human hybrid CcO. Corresponding bacterial pathway has been proposed to convey pumping and water-forming protons, which is supported by mutagenesis. Our analyses did not support the proton pumping function. Study on proton transfer mechanism of proteins by vibrational spectroscopy requires site-specific isotope labeling of proteins, which can be achieved by cell-free protein synthesis. We have successfully developed a E. coli cell free system to produce normal bacterial CcO.
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Report
(4 results)
Research Products
(64 results)