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. 1981 Apr;146(1):385–390. doi: 10.1128/jb.146.1.385-390.1981

Staphylococcus aureus adenosine triphosphatase: inhibitor sensitivity and release from membrane.

B M Kubak, W W Yotis
PMCID: PMC217094  PMID: 6452444

Abstract

Homogeneous preparations of cytoplasmic membrane isolated from Staphylococcus aureus 6538P exhibited membrane-associated adenosine triphosphatase (ATPase) activity. Membrane ATPase activity was activated by divalent cations (4.0 mM: Mg2+ greater than Mn2+ greater than Co2+ greater than Zn2+), and ATP was hydrolyzed more readily than other nucleoside triphosphates and phosphorylated substrates. The pH optimum for the membrane ATPase was 6.5. The ATPase could not be released from the membrane by differential osmotic treatments, but detergent treatment effectively solubilized active enzyme. The nonionic detergent Triton X-100 (1%) released a protein with ATPase activity, after substrate-dependent staining in polyacrylamide gels, that differed slightly in electrophoretic migration when compared to the active enzyme solubilized with sodium dodecyl sulfate (0.1%). Membrane-associated ATPase activity was inhibited by N,N'-dicyclohexylcarbodiimide (0.001 to 1 mM) and NaF (50% inhibition at 5 mM NaF). Azide and trypsin inhibited activity, whereas ouabain had a slight inhibitory effect. Diethylstilbestrol showed appreciable activation of the membrane ATPase over the range employed (0.001 to 1 mM).

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Selected References

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  1. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  2. BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cantley L. C., Jr, Josephson L., Warner R., Yanagisawa M., Lechene C., Guidotti G. Vanadate is a potent (Na,K)-ATPase inhibitor found in ATP derived from muscle. J Biol Chem. 1977 Nov 10;252(21):7421–7423. [PubMed] [Google Scholar]
  4. Collins M. L., Salton M. R. Solubility characteristics of Micrococcus lysodeikticus membrane components in detergents and chaotropic salts analyzed by immunoelectrophoresis. Biochim Biophys Acta. 1979 May 3;553(1):40–53. doi: 10.1016/0005-2736(79)90029-4. [DOI] [PubMed] [Google Scholar]
  5. Epstein F. H., Whittam R. The mode of inhibition by calcium of cell-membrane adenosine-triphosphatase activity. Biochem J. 1966 Apr;99(1):232–238. doi: 10.1042/bj0990232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fillingame R. H. Purification of the carbodiimide-reactive protein component of the ATP energy-transducing system of Escherichia coli. J Biol Chem. 1976 Nov 10;251(21):6630–6637. [PubMed] [Google Scholar]
  7. Fitzgerald T., Yotis W. W. Hormonal intervention in the uptake of amino acids by staphylococci. J Med Microbiol. 1971 Feb;4(1):89–96. doi: 10.1099/00222615-4-1-89. [DOI] [PubMed] [Google Scholar]
  8. Gross R., Coles N. W. Adenosine triphosphatase in isolated membranes of Staphylococcus aureus. J Bacteriol. 1968 Apr;95(4):1322–1326. doi: 10.1128/jb.95.4.1322-1326.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. HURLBERT R. B., SCHMITZ H., BRUMM A. F., POTTER V. R. Nucleotide metabolism. II. Chromatographic separation of acid-soluble nucleotides. J Biol Chem. 1954 Jul;209(1):23–39. [PubMed] [Google Scholar]
  10. Hamilton I. R. Effects of fluoride on enzymatic regulation of bacterial carbohydrate metabolism. Caries Res. 1977;11 (Suppl 1):262–291. doi: 10.1159/000260304. [DOI] [PubMed] [Google Scholar]
  11. Harold F. M., Baarda J. R., Baron C., Abrams A. Inhibition of membrane-bound adenosine triphosphatase and of cation transport in Streptococcus faecalis by N,N'-dicyclohexylcarbodiimide. J Biol Chem. 1969 May 10;244(9):2261–2268. [PubMed] [Google Scholar]
  12. Helenius A., Simons K. Solubilization of membranes by detergents. Biochim Biophys Acta. 1975 Mar 25;415(1):29–79. doi: 10.1016/0304-4157(75)90016-7. [DOI] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Marquis R. E. Inhibition of streptococcal adenosine triphosphatase by fluoride. J Dent Res. 1977 Jun;56(6):704–704. doi: 10.1177/00220345770560063301. [DOI] [PubMed] [Google Scholar]
  15. Muñoz E., Nachbar M. S., Schor M. T., Salton M. R. Adenosinetriphosphatase of Micrococcus lysodeikticus: selective release and relationship to membrane structure. Biochem Biophys Res Commun. 1968 Aug 13;32(3):539–546. doi: 10.1016/0006-291x(68)90696-7. [DOI] [PubMed] [Google Scholar]
  16. Muñoz E., Salton M. R., Ng M. H., Schor M. T. Membrane adenosine triphosphatase of Micrococcus lysodeikticus. Purification, properties of the "soluble" enzyme and properties of the membrane-bound enzyme. Eur J Biochem. 1969 Feb;7(4):490–501. [PubMed] [Google Scholar]
  17. Theodore T. S., Popkin T. J., Cole R. M. The separation and isolation of plasma membranes and mesosomal vesicles from Staphylococcus aureus. Prep Biochem. 1971;1(3):233–248. doi: 10.1080/00327487108081942. [DOI] [PubMed] [Google Scholar]
  18. WINZLER R. J. Determination of serum glycoproteins. Methods Biochem Anal. 1955;2:279–311. doi: 10.1002/9780470110188.ch10. [DOI] [PubMed] [Google Scholar]
  19. Wang C., Smith R. L. Lowry determination of protein in the presence of Triton X-100. Anal Biochem. 1975 Feb;63(2):414–417. doi: 10.1016/0003-2697(75)90363-2. [DOI] [PubMed] [Google Scholar]
  20. Weinbaum G., Markman R. A rapid technique for distinguishing enzymatically active proteins in the cell"envelope" of Escherichia coli B. Biochim Biophys Acta. 1966 Jul 27;124(1):207–209. doi: 10.1016/0304-4165(66)90335-7. [DOI] [PubMed] [Google Scholar]
  21. Yotis W. W., Fitzgerald T. Hormonally induced alterations in S. aureus. Ann N Y Acad Sci. 1974 Jul 31;236(0):187–202. doi: 10.1111/j.1749-6632.1974.tb41491.x. [DOI] [PubMed] [Google Scholar]

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