Cysteine 295 indirectly affects Ni coordination of carbon monoxide dehydrogenase-II C-cluster

Biochem Biophys Res Commun. 2013 Nov 8;441(1):13-7. doi: 10.1016/j.bbrc.2013.09.143. Epub 2013 Oct 10.

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.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehyde Oxidoreductases / chemistry*
  • Aldehyde Oxidoreductases / metabolism*
  • Cysteine / metabolism*
  • Multienzyme Complexes / chemistry*
  • Multienzyme Complexes / metabolism*
  • Multigene Family*
  • Mutant Proteins / chemistry
  • Mutant Proteins / metabolism
  • Nickel / metabolism*
  • Peptococcaceae / enzymology*
  • Phylogeny
  • Sequence Homology, Amino Acid
  • Spectrophotometry, Ultraviolet
  • Structure-Activity Relationship

Substances

  • Multienzyme Complexes
  • Mutant Proteins
  • Nickel
  • Aldehyde Oxidoreductases
  • carbon monoxide dehydrogenase
  • Cysteine