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. 1975 Aug;8(2):146–153. doi: 10.1128/aac.8.2.146

Bactericidal Activity of Superoxide Anion and of Hydrogen Peroxide: Investigations Employing Dialuric Acid, a Superoxide-Generating Drug

Lawrence R DeChatelet *, Pamela S Shirley , Phillip R Goodson , Charles E McCall
PMCID: PMC429281  PMID: 170855

Abstract

The addition of dialuric acid (a superoxide-generating drug) to a suspension of resting human neutrophils resulted in a stimulation of cellular hexose monophosphate shunt activity. Measurement of oxygen consumption demonstrated a rapid rate of oxygen uptake by the drug alone in aqueous solution. The subsequent addition of catalase (but not of superoxide dismutase) resulted in a substantial release of oxygen, indicating that H2O2 was accumulating in the media. The generation of O2 by the drug was verified by measuring the rate of reduction of cytochrome c by the drug in the presence and absence of authentic superoxide dismutase. The growth of Escherichia coli B and Staphylococcus aureus was inhibited in vitro by addition of the drug to a suitable culture media; the degree of inhibition was dose dependent. This inhibition of bacterial growth could be completely reversed by the addition of as little as 5 μg of purified catalase, but was not affected by concentrations of superoxide dismutase that were 2,000-fold higher. These results suggest that the dialuric acid-induced stimulation of hexose monophosphate shunt activity in neutrophils is due to accumulated H2O2. Further, the contribution of superoxide to the bactericidal activity towards E. coli and S. aureus is probably minimal compared with that of H2O2.

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Selected References

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

  1. Allen R. C., Stjernholm R. L., Steele R. H. Evidence for the generation of an electronic excitation state(s) in human polymorphonuclear leukocytes and its participation in bactericidal activity. Biochem Biophys Res Commun. 1972 May 26;47(4):679–684. doi: 10.1016/0006-291x(72)90545-1. [DOI] [PubMed] [Google Scholar]
  2. Allen R. C., Yevich S. J., Orth R. W., Steele R. H. The superoxide anion and singlet molecular oxygen: their role in the microbicidal activity of the polymorphonuclear leukocyte. Biochem Biophys Res Commun. 1974 Oct 8;60(3):909–917. doi: 10.1016/0006-291x(74)90401-x. [DOI] [PubMed] [Google Scholar]
  3. Babior B. M., Kipnes R. S., Curnutte J. T. Biological defense mechanisms. The production by leukocytes of superoxide, a potential bactericidal agent. J Clin Invest. 1973 Mar;52(3):741–744. doi: 10.1172/JCI107236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cohen G., Heikkila R. E. The generation of hydrogen peroxide, superoxide radical, and hydroxyl radical by 6-hydroxydopamine, dialuric acid, and related cytotoxic agents. J Biol Chem. 1974 Apr 25;249(8):2447–2452. [PubMed] [Google Scholar]
  5. Cooper M. R., DeChatelet L. R., McCall C. E., LaVia M. F., Spurr C. L., Baehner R. L. Complete deficiency of leukocyte glucose-6-phosphate dehydrogenase with defective bactericidal activity. J Clin Invest. 1972 Apr;51(4):769–778. doi: 10.1172/JCI106871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dechatelet L. R., Cooper M. R., McCall C. E. Dissociation by colchicine of the hexose monophosphate shunt activation from the bactericidal activity of the leukocyte. Infect Immun. 1971 Jan;3(1):66–72. doi: 10.1128/iai.3.1.66-72.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fee J. A., Teitelbaum H. D. Evidence that superoxide dismutase plays a role in protecting red blood cells against peroxidative hemolysis. Biochem Biophys Res Commun. 1972 Oct 6;49(1):150–158. doi: 10.1016/0006-291x(72)90022-8. [DOI] [PubMed] [Google Scholar]
  8. Gregory E. M., Fridovich I. Oxygen toxicity and the superoxide dismutase. J Bacteriol. 1973 Jun;114(3):1193–1197. doi: 10.1128/jb.114.3.1193-1197.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Holmes B., Page A. R., Good R. A. Studies of the metabolic activity of leukocytes from patients with a genetic abnormality of phagocytic function. J Clin Invest. 1967 Sep;46(9):1422–1432. doi: 10.1172/JCI105634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Karnovsky M. L. The metabolism of leukocytes. Semin Hematol. 1968 Apr;5(2):156–165. [PubMed] [Google Scholar]
  11. Klebanoff S. J. Iodination of bacteria: a bactericidal mechanism. J Exp Med. 1967 Dec 1;126(6):1063–1078. doi: 10.1084/jem.126.6.1063. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Klebanoff S. J. Role of the superoxide anion in the myeloperoxidase-mediated antimicrobial system. J Biol Chem. 1974 Jun 25;249(12):3724–3728. [PubMed] [Google Scholar]
  13. McCord J. M., Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem. 1969 Nov 25;244(22):6049–6055. [PubMed] [Google Scholar]
  14. McRipley R. J., Sbarra A. J. Role of the phagocyte in host-parasite interactions. XI. Relationship between stimulated oxidative metabolism and hydrogen peroxide formation, and intracellular killing. J Bacteriol. 1967 Nov;94(5):1417–1424. doi: 10.1128/jb.94.5.1417-1424.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Misra H. P., Fridovich I. The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem. 1972 May 25;247(10):3170–3175. [PubMed] [Google Scholar]
  16. Reed P. W. Glutathione and the hexose monophosphate shunt in phagocytizing and hydrogen peroxide-treated rat leukocytes. J Biol Chem. 1969 May 10;244(9):2459–2464. [PubMed] [Google Scholar]
  17. 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]
  18. Selvaraj R. J., Sbarra A. J. Relationship of glycolytic and oxidative metabolism to particle entry and destruction in phagocytosing cells. Nature. 1966 Sep 17;211(5055):1272–1276. doi: 10.1038/2111272a0. [DOI] [PubMed] [Google Scholar]
  19. Strauss R. R., Paul B. B., Jacobs A. A., Sbarra A. J. Role of the Phagocyte in Host-Parasite Interactions XXVII. Myeloperoxidase-H(2)O(2)-Cl-Mediated Aldehyde Formation and Its Relationship to Antimicrobial Activity. Infect Immun. 1971 Apr;3(4):595–602. doi: 10.1128/iai.3.4.595-602.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Webb L. S., Keele B. B., Jr, Johnston R. B., Jr Inhibition of phagocytosis-associated chemiluminescence by superoxide dismutase. Infect Immun. 1974 Jun;9(6):1051–1056. doi: 10.1128/iai.9.6.1051-1056.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]

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