Structural Characterization of Subunit-Specific Isotope labelled Membrane Bound Protein

Research Project

Project/Area Number	25650016
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Multi-year Fund
Research Field	Structural biochemistry
Research Institution	Hokkaido University
Principal Investigator	ISHIMORI Koichiro 北海道大学, 理学(系)研究科(研究院), 教授 (20192487)
Co-Investigator(Kenkyū-buntansha)	UCHIDA Takeshi 北海道大学, 大学院理学研究院, 准教授 (30343742)
Project Period (FY)	2013-04-01 – 2015-03-31
Project Status	Completed (Fiscal Year 2014)
Budget Amount *help	¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000) Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000) Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords	シトクロム酸化酵素 / ナノディスク / 呼吸鎖電子伝達系 / NMR / MSP / 安定同位体
Outline of Final Research Achievements	To discuss the electron transfer mechanism in the respiratory chain in mitochondria, the reconstitution of an isotope labelled key enzyme, cytochrome c oxidase, into the nano-disc, a mimic of membrane in cells, has been examined, which allows us to measure the high resolution NMR spectra of the membrane-bound proteins under the physiological condition. The expression, purification and reconstitution of the bacterial cytochrome c oxidase into the nano-disc were confirmed, leading to the high resolution structural analysis of high molecular weight membrane bound proteins.