Synergistic Inhibition of Membrane ATPase and Cell Growth of Helicobacter pylori by ATPase Inhibitors

  • Ki, Mi-Ran (Graduate School of Biotechnology, Korea University) ;
  • Yun, Soon-Kyu (Graduate School of Biotechnology, Korea University) ;
  • Lim, Wang-Jin (Graduate School of Biotechnology, Korea University) ;
  • Hong, Bum-Shik (Graduate School of Biotechnology, Korea University) ;
  • Hwang, Se-Young (Graduate School of Biotechnology, Korea University)
  • Published : 1999.08.01

Abstract

Helicobacter pylori were found to be resistant to azide but sensitive to vanadate, suggesting that defect in the P-type ATPase activity rather than F-type ATPase would be lethal to cell survival or growth. To elucidate the relationship between this enzyme inhibition and H. pylori death, we determined the effect of omeprazole (OMP) plus vanadate on enzyme activity and cell growth. The minimum inhibitory concentration (MIC; ca. 0.8$\mu$mol/disk) of vanadate for H. pylori growth was lowered over l0-fold with the aid of OMP, whereby its inhibitory potential toward the P-type ATPase activity was diametrically increased. Alternatively, we found that this enzyme activity was essential for active transport in H. pylori. From these observations, we strongly suggest that the immediate cause of the growth inhibition of H. pylori cells with OMP and/or vanadate might be defective in the cell's active transport due to the lack of P-type ATPase activity. From the spectral data with circular dichroism (CD) spectroscopy, we found that activated OMP (OAS) at concentration below MIC did not disrupt helical structures of membrane proteins. Separately, we determined the cytopathic effect of OAS by SDS-PAGE, indicating the change in the production of cytoplasmic protein but not cell membrane.

Keywords

References

  1. Gut v.36 Detection of the intragastric sites at which Helicobavter pylori evades treatment with amixicillin and cimetidine Athertonm, J. C.;A. Cockayne;M. Balsitis;G. E. Kirk;C. J. Hawkey;R. C. Spiller
  2. Infect. Immun v.64 Allelic exchange mutagenesis of nixA in Helicobacter pylori results in reduced nickel transport and urease activity Bauerfeind, P.;R. M. Garner;L. T. Mobley
  3. Gut v.40 Effect of N-α-methylhistamine on acid secretionin isolated rabbit rarietal cells: Implications for Helicobacter pylori associated gastritis and gastric physiology Beales, I. L. P.;J. Calam
  4. Pharmacology v.50 Bismuth subcitrate and omeprazole inhibit Helicobacter pylori $F_1$-ATPase Beil. W.;C. Birkholz;S. Wagner;K.-F. Sewing
  5. Antimicrob. Agents Chemother v.39 Partial characterization and effect of omeprazole on ATPase activity in Helicobacter pylori by using permeabilized cells Bell, W. A.;J. Fryklund
  6. J. Biol. Chem. v.272 Sites of reaction of the gastric H, K-ATPase with extracytoplasmic thiol reagents Besancon, M.;A. Simon;G. Sachs;J. M. Shin
  7. Gastroenterology v.93 Gastric Campylobacter-like organisms. Gastritis and peptic ulcer disease Blaser, M. J.
  8. Proc. Natl. Acad. Sci. USA v.85 Bafilomycin: a class of inhibitors of membrane ATPases from microorganisms, animal cell, and plant cells Bowman, E. J.;A. Siebers;K. Altendorf
  9. Microbiology v.140 Characteristics of Helicobacter pylori growth in a defines medium and determination of its amino acid requirements David, J. R.;C. W. Penn
  10. Annu. Rev. Microbiol v.44 Helicobacter (Campylobacter) pylori: A new twist to an old disease Dick, J. D.
  11. Annu. Rev. Biochem v.48 Membrane adenosine triphosphatase of prokaryotic cells Downie, J. A.;F. Gibson;G. B. Cox
  12. Nature (London) v.290 Substituted benzimidazoles inhibit acid secretion by blocking $H^+$, $K^+$ ATPase Fellenius, E.;T. Berglindh;G. Sachs;L. Olbe;B. Elander;S. E. Sjostrand;B. Wallmark
  13. J. clin. Pathol v.39 Campulobacter pyliodis, gastritis and peptic ulceration Goodwin, C. S.
  14. Ann. Intern. Med v.115 Effect of triple therapy (antibiotics plus bismuth) on duodenal ulcer healing Graham, D. Y.;G. W. Lew;D. G. Evans;D. J. Evans;P. D. Klein
  15. Am. J. Med v.100(suppl. 5A) Eradication of Helicobacter pylori infection Hunt, R. H.
  16. Pediatrics v.10 Omeprazole-based dual and triple regimens for Helicobacter pylori eradicationin children Kato, S.;J. Takeyama;K. Ebina;H. Naganuma
  17. Gastroenterology v.116 Helicobacter pylori, acid, and omeprazole revisited: Bavterial eradication and rebound hypersecretion Kuipers, E. J.;E. C. Klinkenberg-Knol
  18. Nature v.227 Cleavage of structural proteins during the assembly of the head protein of bacteriophage T4 Laemmli, U. K.
  19. Gastroenterology v.116 The MACH2 study: Role of omeprazole in eradication fo Helicobacter pylori with 1-week triple therapies Lind, T.;F. Megraud;P. Unge;E. Bayerdorffer;C. O'mrain;R. Spiller;S. V. V. Zanten;K. D. Bardhan;M. Hellblom;M. Wrangstradh;L. Zeijlon;C. Cederberg
  20. Pharmacol. Ther. v.69 The chemotherapeutic effects of $H^+$/$K^+$ ATPase inhibitors on Helicobacter pylori infection Logan, R. P.
  21. Gastroenterology v.107 The proton pump inhibitor omeprazole inhibits acid survival of Helicobacter pylori by a urease-independent mechanism McGowan, C. C.;T. C. Cover;M. J. Blaser
  22. J. Biol. Chem. v.271 Cloning and membrane topology of a P-type ATPase from Helicobacter pylori Melchersm K.;T. Weitzenegger;A. Buhmann;W. Steinhilber;G. Sachs;K. P. Schafer
  23. J. Natl. Cancer Inst. v.89 Infection with Helicobacter pylori $cagA^+$ strains and dissociation of gastric epothelial cell proliferation from apoptosis Peek, R. M.;S. F. Moss;K. T. Tham;G. I. Perez-Pcrez;S. Wang;G. G. Miller;J. C. Atherton;P. R. Holt;M. J. Blaser
  24. Mannual of Clinical Microbiology (5th ed.) Campylobacter, Helicobacter, and related sprial bacteria Penner, J. L.;Balows, A.;W. J. Hausler,Jr.;K. L. Herrmann;H. D. Isenberg;H. J. Shadomy(eds.)
  25. Lancet v.335 Cure of duodenal ulcer associated with eradication of Helicobacter pylori Rauws, E. A. J.;G. N. J. Tytgat
  26. Antimicrob. Agents Chemother v.41 Basis for the selective antibavterial activity in vitro of proton pump inhibitors against Helicobacter spp Sjostrom, J. E.;T. Kuhler;H. Larsson
  27. Eur. J. Clin. Microbiol. Infect. Dis v.10 Antibacterial activity of pantoprazole and omeprazole against Helicobacter pylori Sauerbaum, S.;H. Leying;K. Klem;W. Opferkuch
  28. Lancet v.341 An international association between Helicobacter pylori infection and gastric cancer The Eurogast Study Group
  29. Anal. Biochem v.175 Investigation of the bicinchonic acid protein assay: Identification of the groups responsible for color formation Wiechelman, K.;K. Braun;J. Fitzpatrick
  30. J. Antimicrob. Chemother v.37 Clarithromycin resistance in Helicobacter pylori: Prevalence in untreated dyspeptic patients and stability in vitro Xia, H. X.;M. Buckley;C. T. Leone;C. A. O'Morain
  31. Biochem. Biophys. Res. Commun v.40 On the reversibility of binding of cardiotonic steroids to a partially purified (Na+K)-activated adenosinetriphosphatase from beff brain Yoda, A.;L. E. Hokins
  32. J. Microbiol. Biotechnol v.7 Characteristics of ATPases present in everted membrane vesicle of Helicobacter pylori Yun, S.-K.;S.-Y. Hwang
  33. J. Microbiol. Biotechnol v.77 Expression of latent P-type ATPase and their presumptive roles in cell membrance of Helicobacter pylori Yun, S.-K.;S.-Y. Hwang