| Project/Area Number |
11440198
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Inorganic chemistry
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| Research Institution | Osaka University |
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Principal Investigator |
SUZUKI Shinnichiro Osaka University, Graduate School of Science, Professor, 大学院・理学研究科, 教授 (70116052)
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| Co-Investigator(Kenkyū-buntansha) |
KATAOKA Kunishige Kanazawa University, Faculty of Science, Assistant Professor, 理学部, 講師 (40252712)
YAMAGUCHI Kazuya Osaka University, Graduate School of Science, Assistant Professor, 大学院・理学研究科, 講師 (80252550)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥12,100,000 (Direct Cost: ¥12,100,000)
Fiscal Year 2001: ¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2000: ¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1999: ¥5,300,000 (Direct Cost: ¥5,300,000)
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| Keywords | Multi-nuclear Copper Enzymes / denitrifying bacteria / nitrite reductase / type 1 Cu / type 2 Cu / pulse radiolysis / nitrous oxide reductase / intramolecular electron transfer process / 銅酵素 / タンパク質内電子移動 / 銅タンパク質 / 変異体 / 亜硝酸還元酸素 |
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| Research Abstract |
The purpose of this project is to clarify the structure and function of Cu-containing nitrite reductase and nitrous oxide reductase.
1) In order to shed light on the His255 and Asp98 residues around the type 2 Cu center in blue Achromobacter xylosoxidans nitrite reductase, two mutants were expressed in E. coil.
The Asp98Ala and His255Ala mutants show the very low enzyme activities compared with the wild enzyme at pH 5.5, suggesting that these residues play a role of proton donation for the formation of water molecule.
The Asp and His255 residues are also very important for the control of the intramolecular electron transfer rate from type 1 Cu to type 2.
2) The nitrite reductase from Hyphomicrobium sp. A3151 has the two type 1 Cu's and one type 2 Cu, which is different from the other nitrite reductases containing type 1 Cu : type 2 Cu = 1 : 1. One of type 1 Cu(II) ions is blue and the other is green.
3) The nitrous oxide reductase from Hyphomicrobium sp. A3151 is aerobically stable, and there are two forms, purple oxidized and blue reduced forms.
4) The type 2 Cu(I) and (II) model complexes were prepared. The Cu(I) complex binding nitrite with nitrogen atom stoichiometrically gives NO gas in the presence of proton.
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