title: Characterization of the Type III sulfide:quinone oxidoreductase from Caldivirga maquilingensis and its membrane binding.
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.
authors.affiliation: Department of Biochemistry, University of Illinois, Urbana, IL 61801, USA.
authors.forename: Andrea M
authors.initials: AM
authors.lastname: Lencina
authors.forename: Ziqiao
authors.initials: Z
authors.lastname: Ding
authors.forename: Lici A
authors.initials: LA
authors.lastname: Schurig-Briccio
authors.forename: Robert B
authors.initials: RB
authors.lastname: Gennis
mesh_terms: D000595:Amino Acid Sequence; D001105:Archaea / Q000378:metabolism*; D001665:Binding Sites; D008565:Membrane Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D011808:Quinone Reductases / Q000737:chemistry / Q000378:metabolism*; D013440:Sulfides / Q000378:metabolism*
chemical_list: D008565:Membrane Proteins; D013440:Sulfides; D011808:Quinone Reductases