Chemiosmotic energy conversion and the membrane ATPase of Methanolobus tindarius.
Electron transport phosphorylation has been demonstrated to drive ATP synthesis for the methanogenic archaebacterium Methanolobus tindarius: Protonophores evoked uncoupler effects and lowered the membrane potential delta psi. Under the influence of N,N'-dicyclohexylcarbodiimide [(cHxN)2C] the membrane potential increased while methanol turnover was inhibited. 2-Bromoethanesulfonate, an inhibitor of methanogenesis, had no effect on the membrane potential but, like (cHxN)2C and protonophores, decreased the intracellular ATP concentration. Labeling experiments with (cHxN)2(14)C showed membranes to contain a proteolipid, with a molecular mass of 5.5 kDa, that resembles known (cHxN)2C-binding proteins of F0-F1 ATPases. The (cHxN)2-sensitive membrane ATPase hydrolysed Mg.ATP at a pH optimum of 5.0 with a Km (ATP) of 2.5 mM (V = 77 mU/mg). It was inhibited competitively by ADP; Ki (ADP) = 0.65 mM. Azide or vanadate caused no significant loss in ATPase activity, but millimolar concentrations of nitrate showed an inhibitory effect, suggesting a relationship to ATPases from vacuolar membranes. In contrast, no inhibition occurred in the presence of bafilomycin A1. The ATPase was extractable with EDTA at low salt concentrations. The purified enzyme consists of four different subunits, alpha (67 kDa), beta (52 kDa), gamma (20 kDa) and beta (less than 10 kDa), as determined from SDS gel electrophoresis.
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Institut für Biochemie, Medizinische Universität zu Lübeck, Federal Republic of Germany.
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D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D000255:Adenosine Triphosphate / Q000096:biosynthesis; D001386:Azides / Q000494:pharmacology; D001425:Bacterial Outer Membrane Proteins / Q000302:isolation & purification*; D002462:Cell Membrane / Q000201:enzymology*; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology; D004579:Electron Transport; D015180:Electrophoresis, Gel, Two-Dimensional; D004734:Energy Metabolism; D019605:Euryarchaeota / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D008564:Membrane Potentials; D000432:Methanol / Q000378:metabolism; D009566:Nitrates / Q000494:pharmacology; D010766:Phosphorylation; D012995:Solubility
D001386:Azides; D001425:Bacterial Outer Membrane Proteins; D009566:Nitrates; D004024:Dicyclohexylcarbodiimide; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D000432:Methanol