title: VhuD facilitates electron flow from H2 or formate to heterodisulfide reductase in Methanococcus maripaludis.
abstract: Flavin-based electron bifurcation has recently been characterized as an essential energy conservation mechanism that is utilized by hydrogenotrophic methanogenic Archaea to generate low-potential electrons in an ATP-independent manner. Electron bifurcation likely takes place at the flavin associated with the α subunit of heterodisulfide reductase (HdrA). In Methanococcus maripaludis the electrons for this reaction come from either formate or H2 via formate dehydrogenase (Fdh) or Hdr-associated hydrogenase (Vhu). However, how these enzymes bind to HdrA to deliver electrons is unknown. Here, we present evidence that the δ subunit of hydrogenase (VhuD) is central to the interaction of both enzymes with HdrA. When M. maripaludis is grown under conditions where both Fdh and Vhu are expressed, these enzymes compete for binding to VhuD, which in turn binds to HdrA. Under these conditions, both enzymes are fully functional and are bound to VhuD in substoichiometric quantities. We also show that Fdh copurifies specifically with VhuD in the absence of other hydrogenase subunits. Surprisingly, in the absence of Vhu, growth on hydrogen still occurs; we show that this involves F420-reducing hydrogenase. The data presented here represent an initial characterization of specific protein interactions centered on Hdr in a hydrogenotrophic methanogen that utilizes multiple electron donors for growth.
authors.affiliation: Department of Microbiology, University of Washington, Seattle, Washington, USA.
authors.forename: Kyle C
authors.initials: KC
authors.lastname: Costa
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Lie
authors.forename: Qin
authors.initials: Q
authors.lastname: Xia
authors.forename: John A
authors.initials: JA
authors.lastname: Leigh
mesh_terms: D019843:Archaeal Proteins / Q000378:metabolism*; D004583:Electrons; D004734:Energy Metabolism; D005560:Formate Dehydrogenases / Q000378:metabolism*; D005561:Formates / Q000378:metabolism*; D006859:Hydrogen / Q000378:metabolism*; D017017:Methanococcus / Q000201:enzymology* / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D019843:Archaeal Proteins; D005561:Formates; C432140:VhuD protein, Methanococcus maripaludis; C030544:formic acid; D006859:Hydrogen; D010088:Oxidoreductases; C087171:heterodisulfide reductase; D005560:Formate Dehydrogenases
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