Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1987 Oct;55(10):2518–2525. doi: 10.1128/iai.55.10.2518-2525.1987

Hypochlorous acid-promoted loss of metabolic energy in Escherichia coli.

W C Barrette Jr 1, J M Albrich 1, J K Hurst 1
PMCID: PMC260739  PMID: 2820883

Abstract

Oxidation of Escherichia coli by hypochlorous acid (HOCl) or chloramine (NH2Cl) gives rise to massive hydrolysis of cytosolic nucleotide phosphoanhydride bonds, although no immediate change occurs in either the nucleotide pool size or the concentrations of extracellular end products of AMP catabolism. Titrimetric curves of the extent of hydrolysis coincide with curves for loss of cell viability, e.g., reduction in the adenylate energy charge from 0.8 to 0.1-0.2 accompanies loss of 99% of the bacterial CFU. The oxidative damage caused by HOCl is irreversible within 100 ms of exposure of the organism, although nucleotide phosphate bond hydrolysis requires several minutes to reach completion. Neither HOCl nor NH2Cl reacts directly with nucleotides to hydrolyze phosphoanhydride bonds. Loss of viability is also accompanied by inhibition of induction of beta-galactosidase. The proton motive force, determined from the distribution of 14C-radiolabeled lipophilic ions, declines with incremental addition of HOCl after loss of respiratory function; severalfold more oxidant is required for the dissipation of the proton motive force than for loss of viability. These observations establish a causal link between loss of metabolic energy and cellular death and indicate that the mechanisms of oxidant-induced nucleotide phosphate bond hydrolysis are indirect and that they probably involve damage to the energy-transducing and transport proteins located in the bacterial plasma membrane.

Full text

PDF
2518

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ahmed S., Booth I. R. Quantitative measurements of the proton-motive force and its relation to steady state lactose accumulation in Escherichia coli. Biochem J. 1981 Dec 15;200(3):573–581. doi: 10.1042/bj2000573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albrich J. M., Gilbaugh J. H., 3rd, Callahan K. B., Hurst J. K. Effects of the putative neutrophil-generated toxin, hypochlorous acid, on membrane permeability and transport systems of Escherichia coli. J Clin Invest. 1986 Jul;78(1):177–184. doi: 10.1172/JCI112548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Albrich J. M., Hurst J. K. Oxidative inactivation of Escherichia coli by hypochlorous acid. Rates and differentiation of respiratory from other reaction sites. FEBS Lett. 1982 Jul 19;144(1):157–161. doi: 10.1016/0014-5793(82)80591-7. [DOI] [PubMed] [Google Scholar]
  4. Albrich J. M., McCarthy C. A., Hurst J. K. Biological reactivity of hypochlorous acid: implications for microbicidal mechanisms of leukocyte myeloperoxidase. Proc Natl Acad Sci U S A. 1981 Jan;78(1):210–214. doi: 10.1073/pnas.78.1.210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Atkinson D. E. The energy charge of the adenylate pool as a regulatory parameter. Interaction with feedback modifiers. Biochemistry. 1968 Nov;7(11):4030–4034. doi: 10.1021/bi00851a033. [DOI] [PubMed] [Google Scholar]
  6. Beckerdite S., Mooney C., Weiss J., Franson R., Elsbach P. Early and discrete changes in permeability of Escherichia coli and certain other gram-negative bacteria during killing by granulocytes. J Exp Med. 1974 Aug 1;140(2):396–409. doi: 10.1084/jem.140.2.396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Booth I. R., Mitchell W. J., Hamilton W. A. Quantitative analysis of proton-linked transport systems. The lactose permease of Escherichia coli. Biochem J. 1979 Sep 15;182(3):687–696. doi: 10.1042/bj1820687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Camper A. K., McFeters G. A. Chlorine injury and the enumeration of waterborne coliform bacteria. Appl Environ Microbiol. 1979 Mar;37(3):633–641. doi: 10.1128/aem.37.3.633-641.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chapman A. G., Atkinson D. E. Adenine nucleotide concentrations and turnover rates. Their correlation with biological activity in bacteria and yeast. Adv Microb Physiol. 1977;15:253–306. doi: 10.1016/s0065-2911(08)60318-5. [DOI] [PubMed] [Google Scholar]
  10. Chapman A. G., Fall L., Atkinson D. E. Adenylate energy charge in Escherichia coli during growth and starvation. J Bacteriol. 1971 Dec;108(3):1072–1086. doi: 10.1128/jb.108.3.1072-1086.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dietzler D. N., Leckie M. P., Sternheim W. L., Ungar J. M., Crimmins D. L., Lewis J. W. Regulation of glycogen synthesis and glucose utilization in Escherichia coli during maintenance of the energy charge. Quantitative correlation of changes in the rates of glycogen synthesis and glucose utilization with simultaneous changes in the cellular levels of both glucose 6-phosphate and fructose 1,6-diphosphate. J Biol Chem. 1979 Sep 10;254(17):8276–8287. [PubMed] [Google Scholar]
  12. Elsbach P., Beckerdite S., Pettis P., Franson R. Persistence of regulation of macromolecular synthesis by Escherichia coli during killing by disrupted rabbit granulocytes. Infect Immun. 1974 Apr;9(4):663–668. doi: 10.1128/iai.9.4.663-668.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Elsbach P. On the interaction between phagocytes and micro-organisms. N Engl J Med. 1973 Oct 18;289(16):846–852. doi: 10.1056/NEJM197310182891610. [DOI] [PubMed] [Google Scholar]
  14. Elsbach P., Pettis P., Beckerdite S., Franson R. Effects of phagocytosis by rabbit granulocytes on macromolecular synthesis and degradation in different species of bacteria. J Bacteriol. 1973 Aug;115(2):490–497. doi: 10.1128/jb.115.2.490-497.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Elsbach P., Weiss J. A reevaluation of the roles of the O2-dependent and O2-independent microbicidal systems of phagocytes. Rev Infect Dis. 1983 Sep-Oct;5(5):843–853. doi: 10.1093/clinids/5.5.843. [DOI] [PubMed] [Google Scholar]
  16. Fillingame R. H. The proton-translocating pumps of oxidative phosphorylation. Annu Rev Biochem. 1980;49:1079–1113. doi: 10.1146/annurev.bi.49.070180.005243. [DOI] [PubMed] [Google Scholar]
  17. Foote C. S., Goyne T. E., Lehrer R. I. Assessment of chlorination by human neutrophils. Nature. 1983 Feb 24;301(5902):715–716. doi: 10.1038/301715a0. [DOI] [PubMed] [Google Scholar]
  18. Fox C. F., Kennedy E. P. Specific labeling and partial purification of the M protein, a component of the beta-galactoside transport system of Escherichia coli. Proc Natl Acad Sci U S A. 1965 Sep;54(3):891–899. doi: 10.1073/pnas.54.3.891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Glaser J. H., Conrad H. E. Properties of chick embryo chondrocytes grown in serum-free medium. J Biol Chem. 1984 Jun 10;259(11):6766–6772. [PubMed] [Google Scholar]
  20. Grisham M. B., Jefferson M. M., Melton D. F., Thomas E. L. Chlorination of endogenous amines by isolated neutrophils. Ammonia-dependent bactericidal, cytotoxic, and cytolytic activities of the chloramines. J Biol Chem. 1984 Aug 25;259(16):10404–10413. [PubMed] [Google Scholar]
  21. Hurst J. K., Albrich J. M., Green T. R., Rosen H., Klebanoff S. Myeloperoxidase-dependent fluorescein chlorination by stimulated neutrophils. J Biol Chem. 1984 Apr 25;259(8):4812–4821. [PubMed] [Google Scholar]
  22. KOCH A. L. The inactivation of the transport mechanism for beta-galactosides of Escherichia coli under various physiological conditions. Ann N Y Acad Sci. 1963 Jan 21;102:602–620. doi: 10.1111/j.1749-6632.1963.tb13663.x. [DOI] [PubMed] [Google Scholar]
  23. Kashket E. R. Effects of aerobiosis and nitrogen source on the proton motive force in growing Escherichia coli and Klebsiella pneumoniae cells. J Bacteriol. 1981 Apr;146(1):377–384. doi: 10.1128/jb.146.1.377-384.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Leung H. B., Schramm V. L. Adenylate degradation in Escherichia coli. The role of AMP nucleosidase and properties of the purified enzyme. J Biol Chem. 1980 Nov 25;255(22):10867–10874. [PubMed] [Google Scholar]
  25. Niven D. F., Collins P. A., Knowles C. J. Adenylate energy charge during batch culture of Beneckea natriegens. J Gen Microbiol. 1977 Jan;98(1):95–108. doi: 10.1099/00221287-98-1-95. [DOI] [PubMed] [Google Scholar]
  26. Paul B. B., Jacobs A. A., Strauss R. R., Sbarra A. J. Role of the Phagocyte in Host-Parasite Interactions XXIV. Aldehyde Generation by the Myeloperoxidase-H(2)O(2)-Chloride Antimicrobial System: a Possible In Vivo Mechanism of Action. Infect Immun. 1970 Oct;2(4):414–418. doi: 10.1128/iai.2.4.414-418.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rottenberg H. The measurement of membrane potential and deltapH in cells, organelles, and vesicles. Methods Enzymol. 1979;55:547–569. doi: 10.1016/0076-6879(79)55066-6. [DOI] [PubMed] [Google Scholar]
  28. Schweinsberg P. D., Loo T. L. Simultaneous analysis of ATP, ADP, AMP, and other purines in human erythrocytes by high-performance liquid chromatography. J Chromatogr. 1980 Jan 11;181(1):103–107. doi: 10.1016/s0378-4347(00)81276-1. [DOI] [PubMed] [Google Scholar]
  29. Selsted M. E., Szklarek D., Lehrer R. I. Purification and antibacterial activity of antimicrobial peptides of rabbit granulocytes. Infect Immun. 1984 Jul;45(1):150–154. doi: 10.1128/iai.45.1.150-154.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Selvaraj R. J., Paul B. B., Strauss R. R., Jacobs A. A., Sbarra A. J. Oxidative peptide cleavage and decarboxylation by the MPO-H2O2-Cl- antimicrobial system. Infect Immun. 1974 Feb;9(2):255–260. doi: 10.1128/iai.9.2.255-260.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sips H. J., Hamers M. N. Mechanism of the bactericidal action of myeloperoxidase: increased permeability of the Escherichia coli cell envelope. Infect Immun. 1981 Jan;31(1):11–16. doi: 10.1128/iai.31.1.11-16.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Swedes J. S., Sedo R. J., Atkinson D. E. Relation of growth and protein synthesis to the adenylate energy charge in an adenine-requiring mutant of Escherichia coli. J Biol Chem. 1975 Sep 10;250(17):6930–6938. [PubMed] [Google Scholar]
  33. Taylor M. W., Hershey H. V., Levine R. A., Coy K., Olivelle S. Improved method of resolving nucleotides by reversed-phase high-performance liquid chromatography. J Chromatogr. 1981 Nov 27;219(1):133–139. doi: 10.1016/s0021-9673(00)80584-1. [DOI] [PubMed] [Google Scholar]
  34. Thomas E. L., Grisham M. B., Melton D. F., Jefferson M. M. Evidence for a role of taurine in the in vitro oxidative toxicity of neutrophils toward erythrocytes. J Biol Chem. 1985 Mar 25;260(6):3321–3329. [PubMed] [Google Scholar]
  35. Thomas E. L. Myeloperoxidase, hydrogen peroxide, chloride antimicrobial system: nitrogen-chlorine derivatives of bacterial components in bactericidal action against Escherichia coli. Infect Immun. 1979 Feb;23(2):522–531. doi: 10.1128/iai.23.2.522-531.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Thomas E. L. Myeloperoxidase-hydrogen peroxide-chloride antimicrobial system: effect of exogenous amines on antibacterial action against Escherichia coli. Infect Immun. 1979 Jul;25(1):110–116. doi: 10.1128/iai.25.1.110-116.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Weiss S. J., Klein R., Slivka A., Wei M. Chlorination of taurine by human neutrophils. Evidence for hypochlorous acid generation. J Clin Invest. 1982 Sep;70(3):598–607. doi: 10.1172/JCI110652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Zgliczyński J. M., Stelmaszyńska T. Chlorinating ability of human phagocytosing leucocytes. Eur J Biochem. 1975 Aug 1;56(1):157–162. doi: 10.1111/j.1432-1033.1975.tb02218.x. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES