Structural Study of the Mechanism of Hydrogen Activation on Ni-enzymes
| Project/Area Number |
22370061
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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 |
Biophysics
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| Research Institution | University of Hyogo |
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Principal Investigator |
HIGUCHI Yoshiki 兵庫県立大学, 大学院・生命理学研究科, 教授 (90183574)
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| Co-Investigator(Renkei-kenkyūsha) |
SHOMURA Yasuhito 兵庫県立大学, 大学院・生命理学研究科, 助教 (50423900)
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| Project Period (FY) |
2010 – 2012
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| Project Status |
Completed (Fiscal Year 2012)
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| Budget Amount *help |
¥18,330,000 (Direct Cost: ¥14,100,000、Indirect Cost: ¥4,230,000)
Fiscal Year 2012: ¥7,670,000 (Direct Cost: ¥5,900,000、Indirect Cost: ¥1,770,000)
Fiscal Year 2011: ¥7,670,000 (Direct Cost: ¥5,900,000、Indirect Cost: ¥1,770,000)
Fiscal Year 2010: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
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| Keywords | ヒドロゲナーゼ / X線結晶解析 / 反応機構 / Ni酵素 / 燃料電池 |
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| Research Abstract |
We have studied newly found NAD+-reducing four subunits [NiFe] hydrogenase and O_2-tolerant membrane-bound [NiFe] hydrogenase (MBH) in this project. In 2010-2012, we have developed the new protocols for inoculation of the bacterium and purification for NAD+-reducing four subunits [NiFe] hydrogenase. For MBH, the x-ray crystal structures of as-isolated, H2-reduced and chemically super-oxidized MBH at high resolution has been successfully determined. Overall structure of the molecule and the coordination geometry of the Dinuclear Ni-Fe active site of the MBH are almost identical to those of the standard O_2-sensitive [NiFe] hydrogenase. The proximal iron-sulfur (Fe-S) cluster, however, has a [4Fe-3S]-6Cys structure. When the enzyme is chemically oxidized, the cluster supplies additional electrons to the Ni-Fe active site, and a deprotonated amide nitrogen of the polypeptide backbone coordinates the Fe atom stabilizing the super-oxidized state of the cluster.
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Report
(4 results)
Research Products
(110 results)
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[Journal Article] Studies on hydrogenase2013
Author(s)
T. Yagi, Y. Higuchi
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Journal Title
Proceedings of the Japan Academy, Series B
Volume: 89
Issue: 1
Pages: 16-33
DOI
NAID
ISSN
0386-2208, 1349-2896
Related Report
Peer Reviewed
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[Journal Article] Cytochrome c polymerization by successive domain swapping at the C-terminal helix2010
Author(s)
S. Hirota, Y. Hattori, S. Nagao, M. Taketa, H. Komori, H. Kamikubo, Z. Wang, I. Takahashi, S. Negi, Y. Sugiura, M. Kataoka, Y. Higuchi
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Journal Title
Pro. Natl. Acad. Sci
Volume: 107(29)
Issue: 29
Pages: 12854-12859
DOI
NAID
Related Report
Peer Reviewed
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[Journal Article] Crystal Structures of Ethanolamine Ammonia-lyase complexed with Coenzyme B_12 Analogs and Substrates2010
Author(s)
N. Shibata, H. Tamagaki, N. Hieda, K. Akita, H. Komori, Y. Shomura, S. Terawaki, K. Mori, N. Yasuoka, Y. Higuchi, and T. Toraya
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Journal Title
J. Biol. Chem
Volume: 285 (34)
Issue: 34
Pages: 26484-26493
DOI
Related Report
Peer Reviewed
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[Journal Article] X-ray Crystallographic Analysis of the 6-aminohexanoate Cyclic Dimer Hydrolase: Catalytic Mechanism and Evolution of an Enzyme Responsible for Nylon-6 Byproduct Degradation2010
Author(s)
K. Yasuhira, N. Shibata, G. Mongami, Y. Uedo, Y. Atsumi, Y. Kawashima, A. Hibino, Y. Tanaka, Y-H. Lee, D. Kato, M. Takeo, Y. Higuchi, and S. Negoro
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Journal Title
J. Biol. Chem
Volume: 285
Issue: 2
Pages: 1239-1248
DOI
Related Report
Peer Reviewed
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[Journal Article] Expression, Crystallization and Preliminary X-ray Crystallographic Study of Ethanolamine Ammonia-lyase from Escherichia coli2010
Author(s)
N. Shibata, H. Tamagaki, S. Ohtsuki, N. Hieda, K. Akita, H. Komori, Y. Shomura, S. Terawaki, T. Toraya, N. Yasuoka and Y. Higuchi
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Journal Title
Acta Crystallogr.
Volume: F66
Issue: 6
Pages: 709-711
DOI
Related Report
Peer Reviewed
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[Presentation] Polymerization and Peroxidase Activity Enhancement of Cytochrome c by Successive Domain Swapping at the C-Terminal Helix2012
Author(s)
S. Hirota, Y. Hattori, Z. Wang, S. Nagao, M. Taketa, H. Komori, H. Kamikubo, T. Matsuo, I. Takahashi, S. Negi, Y. Sugiura, M. Kataoka, Y. Higuchi
Organizer
IEEE International Conference on Computational Photography
Place of Presentation
Seattle (USA)
Related Report
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[Presentation] Polymerization and Peroxidase Activity Enhancement of Cytochrome c by Successive Domain Swapping at the C-Terminal Helix2012
Author(s)
S. Hirota, Y. Hattori, Z. Wang, S. Nagao, M. Taketa, H. Komori, H. Kamikubo, T. Matsuo, I. Takahashi, S. Negi, Y. Sugiura, M. Kataoka, Y. Higuchi
Organizer
International Conference on Porphyrins and Phthalocyanins
Place of Presentation
Pusan (Korea)
Related Report
Invited
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