title: H2: heterodisulfide oxidoreductase complex from Methanobacterium thermoautotrophicum. Composition and properties.
abstract: The reduction of the heterodisulfide (CoM-S-S-HTP) of coenzyme M (H-S-CoM) and N-7-mercaptoheptanoylthreonine phosphate (H-S-HTP) with H2 is an energy-conserving step in most methanogenic Archaea. In this study, we show that in Methanobacterium thermoautotrophicum (strain Marburg) this reaction is catalyzed by a stable H2-heterodisulfide oxidoreductase complex of F420-non-reducing hydrogenase and heterodisulfide reductase. This complex, which was loosely associated with the cytoplasmic membrane, was purified 17-fold with 80% yield to apparent homogeneity. The purified complex was composed of six different subunits of apparent molecular masses 80, 51, 41, 36, 21 and 17 kDa, and 1 mol complex, with apparent molecular mass 250 kDa, contained approximately 0.6 mol nickel, 0.9 mol FAD, 26 mol non-heme iron and 22 mol acid-labile sulfur. In 25 mM Chaps, the complex partially dissociated into two subcomplexes. The first subcomplex was was composed of the 51-, 41- and 17-kDa subunits; 1 mol trimer contained 0.7 mol nickel, 10 mol non-heme iron and 9 mol acid-labile sulfur and exhibited F420-non-reducing hydrogenase activity. The other subcomplex was composed of the 80-, 36- and 21-kDa subunits; 1 mol trimer contained 0.8 mol FAD, 22 mol non-heme iron and 15 mol acid-labile sulfur and exhibited heterodi-sulfide-reductase activity. The stimulatory effects of potassium phosphate, a membrane component, uracil derivatives and coenzyme F430 on the H2:heterodisulfide-oxidoreductase activity of the purified complex are described.
authors.affiliation: Max-Planck-Institut für Terrestrsche Mikrobiologie, Marburg, Germany.
authors.forename: E
authors.initials: E
authors.lastname: Setzke
authors.forename: R
authors.initials: R
authors.lastname: Hedderich
authors.forename: S
authors.initials: S
authors.lastname: Heiden
authors.forename: R K
authors.initials: RK
authors.lastname: Thauer
mesh_terms: D002384:Catalysis; D008565:Membrane Proteins / Q000378:metabolism; D017014:Methanobacterium / Q000201:enzymology* / Q000378:metabolism; D008970:Molecular Weight; D010088:Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D011487:Protein Conformation; D012995:Solubility
chemical_list: D008565:Membrane Proteins; D010088:Oxidoreductases; C066352:coenzyme M-7-merceptoheptanoylthreonine-phosphate-heterodisulfide hydrogenase