| 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)
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| 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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