Isolation and characterization of an uncoupler-resistant mutant of Methanothermobacter thermautotrophicus.
A spontaneous mutant of Methanothermobacter thermautotrophicus resistant to the protonophorous uncoupler TCS was isolated. The mutant strain exhibited increased CH(4) formation and elevated level of ATPase activity under non-growing conditions. ATP synthesis driven by methanogenic electron transport as well as by potassium diffusion potential in the presence of either H(+) or Na(+) ions was markedly diminished in the mutant strain. An abundant membrane-associated protein complex with molecular mass approximately 670 kDa was detected in the mutant strain after native PAGE. The results indicate that TCS resistance in this mutant has arisen as a consequence of mutation(s) that affects a specific locus coding for an uncoupler binding protein(s) and/or modulate the activity of unidentified ATPase.
Cubonová
L'ubomíra
L
Institute of Animal Biochemistry and Genetics, Slovak Academy of Sciences, 900 28 Ivanka pri Dunaji, Slovak Republic.
Surín
Stanislav
S
Majerník
Alan
A
Smigán
Peter
P
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000032:analysis / Q000096:biosynthesis; D001426:Bacterial Proteins / Q000032:analysis; D002352:Carrier Proteins / Q000235:genetics / Q000502:physiology; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D004734:Energy Metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006859:Hydrogen / Q000378:metabolism; D008565:Membrane Proteins / Q000032:analysis; D008697:Methane / Q000378:metabolism; D017013:Methanobacteriaceae / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D009154:Mutation; D011188:Potassium / Q000378:metabolism; D012458:Salicylanilides / Q000494:pharmacology*; D012964:Sodium / Q000378:metabolism; D014475:Uncoupling Agents / Q000494:pharmacology*
D001426:Bacterial Proteins; D002352:Carrier Proteins; D004791:Enzyme Inhibitors; D008565:Membrane Proteins; D012458:Salicylanilides; D014475:Uncoupling Agents; C030559:uncoupler-binding protein, Bos taurus; D006859:Hydrogen; D000255:Adenosine Triphosphate; D012964:Sodium; D000251:Adenosine Triphosphatases; C003390:3,3',4',5-tetrachlorosalicylanilide; D008697:Methane; D011188:Potassium