Purification and properties of an ATPase from Sulfolobus solfataricus.
A sulfite-activated ATPase isolated from Sulfolobus solfataricus had a relative molecular mass of 370,000. It was composed of three subunits whose relative molecular masses were 63,000, 48,000, and 24,000. The enzyme was inhibited by the vacuolar ATPase inhibitors nitrate and N-ethylmaleimide; 4-chloro-7-nitrobenzofurazan (NBD-Cl) was also inhibitory. N-Ethylmaleimide was predominately bound to the largest subunit while NBD-Cl was bound to both subunits. ATPase activity was inhibited by low concentrations of p-chloromercuriphenyl sulfonate and the inhibition was reversed by cysteine which suggested that thiol groups were essential for activity. While the ATPase from S. solfataricus shared several properties with the ATPase from S. acidocaldarius there were significant differences. The latter enzyme was activated by sulfate and chloride and was unaffected by N-ethylmaleimide, whereas the S. solfataricus ATPase was inhibited by these anions as well as N-ethylmaleimide. These differences as well as differences that occur in other vacuolar-like ATPases isolated from the methanogenic and the extremely halophilic bacteria suggest the existence of several types of archaeal ATPases, none of which have been demonstrated to synthesize ATP.
Hochstein
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Ames Research Center, Moffett Field, California 94035.
Stan-Lotter
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D009327:4-Chloro-7-nitrobenzofurazan / Q000494:pharmacology; D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D000838:Anions / Q000494:pharmacology; D002729:Chloromercuribenzoates / Q000494:pharmacology; D003545:Cysteine / Q000494:pharmacology; D004789:Enzyme Activation; D005033:Ethylmaleimide / Q000494:pharmacology; D007425:Intracellular Membranes / Q000201:enzymology; D011487:Protein Conformation; D013447:Sulfites / Q000494:pharmacology; D016944:Sulfolobus / Q000201:enzymology*; D014617:Vacuoles / Q000201:enzymology
D000838:Anions; D002729:Chloromercuribenzoates; D013447:Sulfites; D000251:Adenosine Triphosphatases; D009327:4-Chloro-7-nitrobenzofurazan; D003545:Cysteine; D005033:Ethylmaleimide