title: Cysteine 295 indirectly affects Ni coordination of carbon monoxide dehydrogenase-II C-cluster.
abstract: A unique [Ni-Fe-S] cluster (C-cluster) constitutes the active center of Ni-containing carbon monoxide dehydrogenases (CODHs). His(261), which coordinates one of the Fe atoms with Cys(295), is suggested to be the only residue required for Ni coordination in the C-cluster. To evaluate the role of Cys(295), we constructed CODH-II variants. Ala substitution for the Cys(295) substitution resulted in the decrease of Ni content and didn't result in major change of Fe content. In addition, the substitution had no effect on the ability to assemble a full complement of [Fe-S] clusters. This strongly suggests Cys(295) indirectly and His(261) together affect Ni-coordination in the C-cluster.
authors.affiliation: Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Inoue
authors.forename: Kyosuke
authors.initials: K
authors.lastname: Takao
authors.forename: Takashi
authors.initials: T
authors.lastname: Yoshida
authors.forename: Kei
authors.initials: K
authors.lastname: Wada
authors.forename: Takashi
authors.initials: T
authors.lastname: Daifuku
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Yoneda
authors.forename: Keiichi
authors.initials: K
authors.lastname: Fukuyama
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Sako
mesh_terms: D000445:Aldehyde Oxidoreductases / Q000737:chemistry* / Q000378:metabolism*; D003545:Cysteine / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000378:metabolism*; D005810:Multigene Family; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D009532:Nickel / Q000378:metabolism*; D010459:Peptococcaceae / Q000201:enzymology*; D010802:Phylogeny; D017386:Sequence Homology, Amino Acid; D013056:Spectrophotometry, Ultraviolet; D013329:Structure-Activity Relationship
chemical_list: D009097:Multienzyme Complexes; D050505:Mutant Proteins; D009532:Nickel; D000445:Aldehyde Oxidoreductases; C019823:carbon monoxide dehydrogenase; D003545:Cysteine
keywords: C-cluster; CODH; Carbon monoxide dehydrogenase; Carboxydothermus hydrogenoformans; CoA; IPTG; NH(2)OH; PCR; [Ni–Fe–S] cluster; coenzyme; hydroxylamine; isopropyl β-d-1-thiogalactopyranoside; polymerase chain reaction