ATP synthesis at 100 degrees C by an ATPase purified from the hyperthermophilic archaeon Pyrodictium abyssi.
The chemolithoautotrophic archaeon Pyrodictium abyssi isolate TAG 11 lives close to 100 degrees C and gains energy by sulfur respiration, with hydrogen as electron donor. From the membranes of this hyperthermophile, an ATPase complex was isolated. The purified enzyme consists of six major polypeptides, the 67, 51, 41, 26 and 22 kDa subunits composing the AF(1) headpiece, and the 7 kDa proteolipid of the AF(0) component. The headpiece of the enzyme restored the formation of ATP during sulfur respiration in membrane vesicles from which it had been removed by low salt treatment. Characteristics of the reconstituted activity suggest that the same enzyme is responsible for ATP formation in untreated membranes. ATP formation was neither sensitive to ionophores and uncouplers, nor to dicyclohexyl carbodiimide, but depended on closed vesicles. Both ATPase activity (up to 2 micromol per min and mg protein) as well as ATP formation (up to 0.4 micromol per min and mg membrane protein) were highest at 100 degrees C. A P/e2 ratio of close to one can be estimated for sulfur respiration with hydrogen. In addition to ATP, autoradiographic detection revealed the formation of high quantities of (33)P(i)-labeled ADP and of another compound not identified so far.
Dirmeier
R
R
Universität Regensburg, Lehrstuhl für Mikrobiologie, Universitätsstrasse 31, 93053, Regensburg, Germany.
Hauska
G
G
Stetter
K O
KO
D000244:Adenosine Diphosphate / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism* / Q000494:pharmacology; D000595:Amino Acid Sequence; D002413:Cations, Divalent / Q000494:pharmacology; D019069:Cell Respiration / Q000187:drug effects; D019620:Desulfurococcaceae / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology; D006358:Hot Temperature; D006859:Hydrogen / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis / Q000187:drug effects; D007425:Intracellular Membranes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D007476:Ionophores / Q000494:pharmacology; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010710:Phosphates / Q000378:metabolism; D013447:Sulfites / Q000494:pharmacology; D013455:Sulfur / Q000378:metabolism; D014475:Uncoupling Agents / Q000494:pharmacology
D002413:Cations, Divalent; D007476:Ionophores; D010710:Phosphates; D013447:Sulfites; D014475:Uncoupling Agents; D004024:Dicyclohexylcarbodiimide; D000244:Adenosine Diphosphate; D013455:Sulfur; D006859:Hydrogen; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases