Characterization of the Type III sulfide:quinone oxidoreductase from Caldivirga maquilingensis and its membrane binding

Andrea M. Lencina, Ziqiao Ding, Lici A. Schurig-Briccio, Robert B Gennis

Research output: Contribution to journalArticle

Abstract

Sulfide:quinone oxidoreductases (SQRs) are ubiquitous enzymes which have multiple roles: sulfide detoxification, energy generation by providing electrons to respiratory or photosynthetic electron transfer chains, and sulfide homeostasis. A recent structure-based classification defines 6 groups of putative SQRs (I-VI), and representatives of all but group III have been confirmed to have sulfide oxidase activity. In the current work, we report the first characterization of a predicted group III SQR from Caldivirga maquilingensis, and confirm that this protein is a sulfide oxidase. The gene encoding the enzyme was cloned, and the protein was expressed in E. coli and purified. The enzyme oxidizes sulfide using decylubiquinone as an electron acceptor, and is inhibited by aurachin C and iodoacetamide. Analysis of the amino acid sequence indicates that the C. maquilingensis SQR has two amphiphilic helices at the C-terminus but lacks any transmembrane helices. This suggests that C. maquilingensis SQR interacts with the membrane surface and that the interactions are mediated by the C-terminal amphiphilic helices. Mutations within the last C-terminal amphiphilic helix resulted in a water-soluble form of the enzyme which, remarkably, retains full SQR activity using decylubiquinone as the electron acceptor. Mutations at one position, L379, also located in the C-terminal amphiphilic helix, inactivated the enzyme by preventing the interaction with decylubiquinone. It is concluded that the C-terminal amphiphilic helix is important for membrane binding and for forming part of the pathway providing access of the quinone substrate to the protein-bound flavin at the enzyme active site.

Original languageEnglish (US)
Pages (from-to)266-275
Number of pages10
JournalBiochimica et Biophysica Acta - Bioenergetics
Volume1827
Issue number3
DOIs
StatePublished - Mar 1 2013

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Keywords

  • Amphiphilic helix
  • Caldivirga maquilingensis
  • Monotopic membrane protein
  • Sulfide:quinone oxidoreductase

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology

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