| Project/Area Number |
62470144
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
結晶学
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| Research Institution | Himeji Institute of Technology |
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Principal Investigator |
YASUOKA Noritake Professor, Basic Research Laboratory, Himeji Institute of Technology, 工学基礎研究所, 教授 (40029054)
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| Co-Investigator(Kenkyū-buntansha) |
MORIMOTO Yukio Assistant,Basic Research Laboratory,Himeji Institute of Technology, 工学基礎研究所, 助手 (80200450)
HIGUCHI Yoshiki Lecturer,Basic Research Laboratory,Himeji Institute of Technology, 工学基礎研究所, 講師 (90183574)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥5,600,000 (Direct Cost: ¥5,600,000)
Fiscal Year 1988: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1987: ¥4,100,000 (Direct Cost: ¥4,100,000)
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| Keywords | Sulfate Reducin Bacteria / High Molecular Weight Cytochrome C / X-Ray Analysis / チトクロムC553 / チトクロムC_<553> / イメージングプレート |
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| Research Abstract |
1. High molecular weight cytochrome c from desulfovibrio vulgaris hildenborough has been isolated, purified and crystallized. The molecular weight was estimated to be 75,000 as the mean value from the results of gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis. The protein contains 16 c-type heme groups per molecule. The purified high molecular weight cytochrome c shows typical c-type cytochrome absorption spectra, with an -peak at 553.2 nm in the ferroform. The amino acid composition shows that the protein contains a sufficient number of cysteine and histidine residue to account for the high content of heme groups. The amino acid composition of D. vulgaris Hildenborough high molecular weight cytochrome c has no similarities to the D. desulfuricans hexahemoprotein nitrate reductase. The high content of the heme groups might suggest that the cytochrome has an alternate intrinsjc biological function. Crystals of the high molecular weight cytochrome c have been
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grown from solutions of poly (ethylene glycol) 1000 or 2-methyl-2,4-pentanediol. The crystals are in space group P6_2 pr P6_4 with unit cell dimensions a=b=227.8, c=105.7 A and =120 . A large unit cell volume of 4.75・10^6 A^3 suggests that there are four or five protein molecules per asymmetric unit. They diffract to better than 4 A resolution and appear to be resistent to radiation damage. 2. The hydrogenase solubilized from the particulate fraction from Desulfovibrio vulgaris Miyazaki F has been crystallized. The apparently homogeneous hydrogenase has been separated into three components of similar molecular weight by high performance liquid chromatography on DEAE-Toyopearl. Each hydrogenase components was successfully crystallized by means of the vapour diffusion method with polyethylene glycol or 2-methyl-2,4-pentanediol as a precipitating agent. The crystal data: space group P212121, unit cell dimensions, a=102.1, b=126.8, c=66.9 A. Multiwavelenght anomalous dispersion method was applied to a structure analysis of hydrogenase and resulted to give plasuible irosulfur clusters. 3. Structure analysis of cytochrome c553 has been succeeded. Less
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