title: The ATPase of Bacillus alcalophilus. Purification and properties of the enzyme.
abstract: The ATPase of Bacillus alcalophilus was extracted from the bacterial membranes with Triton X-100 and purified by hydroxyapatite column chromatography. SDS gel-electrophoresis of the purified protein indicated the typical subunit pattern of an F1F0 structure with five F1 subunits (alpha, beta, gamma, delta, epsilon) and three F0 subunits (a,b,c). The alpha and beta subunits were antigens for an antiserum against the corresponding subunits of the ATPase of Escherichia coli. Subunit c was extracted from the bacterial membranes with chloroform/methanol. Its amino acid composition was in the range of subunits c from other ATPases. Maximal ATPase activity was observed in the presence of 2-5 mM MgCl2, an ATP/Mg2+ ratio of 2:1 and 25% methanol. In the absence of methanol, only about 1% of the maximal activity was observed. The enzyme was also activated by Ca2+ (in the absence of methanol), reaching about 30% of the maximal activity. The dependence of initial velocity versus ATP of the Ca2(+)-activated but not of the Mg2+/methanol-activated enzyme indicted cooperativity with three strongly cooperative binding sites.
authors: Mikrobiologisches Institut der Eidgenössischen Technischen Hochschule, Zürich, Switzerland.
authors: A
authors: A
authors: Hoffmann
authors: P
authors: P
authors: Dimroth
mesh_terms: D000251:Adenosine Triphosphatases / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D000596:Amino Acids / Q000737:chemistry; D001407:Bacillus / Q000201:enzymology* / Q000254:growth & development; D002118:Calcium / Q000494:pharmacology; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D000432:Methanol / Q000494:pharmacology; D009947:Organotin Compounds / Q000494:pharmacology
chemical_list: D000596:Amino Acids; D009947:Organotin Compounds; D000251:Adenosine Triphosphatases; D008274:Magnesium; D002118:Calcium; D000432:Methanol