2012 Fiscal Year Final Research Report
De novo design of binuclear metalloproteins
| Project/Area Number |
22550150
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Chemistry related to living body
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| Research Institution | Nagoya Institute of Technology |
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Principal Investigator |
TANAKA Toshiki 名古屋工業大学, 工学研究科, 教授 (70171775)
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| Project Period (FY) |
2010 – 2012
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| Keywords | タンパク質工学 |
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| Research Abstract |
The CuA site in cytochrome C oxidase (COX) contains a binuclear copper center, which is composed of two bridging Cys residues forming a Cu_2S_2 diamond cluster. We designed this copper configuration in the de novo designed 4-helix bundled protein. The designed protein exhibited the same UV vis and ESR spectra as those of COX, indicating that we successfully constructed the Cu_2S_2 diamond cluster in the 4-helix bundled protein. Next, we designed a metal configuration from Zn-Cu superoxide dismutase or hemocyanin. We placed six His residues in the designed protein. The designed protein bound one Cu ion. Then, we added Cys residue to monitor the metal binding. We obtained an evidence that shows this mutant protein might bind two Cu ions or Cu and Zn ions simultaneously. The Cys residue might determine the position of Cu ion in the protein. A new strategy to design the plural metal bindings can be proposed using one Cys residue. The detail analyses are required, we are able to construct the metal binding following the new design method.
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Research Products
(10 results)
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[Journal Article] Creation of a binuclear purple copper site within a de novo coiled-coil protein.2012
Author(s)
D.Shiga, Y. Funahashi, H. Masuda, A.Kikuchi, M. Noda, S. Uchiyama, K. Fukui,K. Kanaori, K. Tajima, Y. Takano, H.Nakamura, M. Kamei, T. Tanaka
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Journal Title
Biochemistry
Volume: vol.51
Pages: 7901-7907
Peer Reviewed
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