論文

査読有り
2006年2月

Redox interaction of cytochrome c(3) with [NiFe] hydrogenase from Desulfovibrio vulgaris Miyazaki F

BIOCHEMISTRY
  • N Yahata
  • ,
  • T Saitoh
  • ,
  • Y Takayama
  • ,
  • K Ozawa
  • ,
  • H Ogata
  • ,
  • Y Higuchi
  • ,
  • H Akutsu

45
6
開始ページ
1653
終了ページ
1662
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1021/bi0514360
出版者・発行元
AMER CHEMICAL SOC

Cytochrome (C3) isolated from a sulfate-reducing bacterium, Desulfovibrio vulgaris Miyazaki F, is a tetraheme protein. Its physiological partner, [NiFe] hydrogenase, catalyzes the reversible oxidoreduction of molecular hydrogen. To elucidate the mechanism of electron transfer between cytochrome C3 and [NiFe] hydrogenase, the transient complex formation by these proteins was investigated by means of NMR. All NH signals of uniformly N-15-labeled ferric cytochrome C3 except N-terminus, Pro, and Gly73 were assigned. H-1-N-15 HSQC spectra were recorded for 15N-labeled ferric and ferrous cytochrome C3, in the absence and presence of hydrogenase. Chemical shift perturbations were observed in the region around heme 4 in both oxidation states. Additionally, the region between hemes I and 3 in ferrous cytochrome C3 was affected in the presence of hydrogenase, suggesting that the mode of interaction is different in each redox state. Heme 3 is probably the electron gate for ferrous cytochrome C3. To investigate the transient complex of cytochrome C3 and hydrogenase in detail, modeling of the complex was performed for the oxidized proteins using a docking program, ZDOCK 2.3, and NMR data. Furthermore, the roles of lysine residues of cytochrome C3 in the interaction with hydrogenase were investigated by site-directed mutagenesis. When the lysine residues around heme 4 were replaced by an uncharged residue, methionine, one by one, the, K-m, of the electron-transfer kinetics increased. The results showed that the positive charges of Lys60, Lys72, Lys95, and Lys101 around heme 4 are important for formation of the transient complex with [NiFe] hydrogenase in the initial stage of the cytochrome C3 reduction. This finding is consistent with the most possible structure of the transient complex obtained by modeling.

Web of Science ® 被引用回数 : 34

リンク情報
DOI
https://doi.org/10.1021/bi0514360
CiNii Articles
http://ci.nii.ac.jp/naid/30010865741
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/16460012
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000235631400013&DestApp=WOS_CPL
ID情報
  • DOI : 10.1021/bi0514360
  • ISSN : 0006-2960
  • CiNii Articles ID : 30010865741
  • PubMed ID : 16460012
  • Web of Science ID : WOS:000235631400013

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