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. 1986 Jan 15;233(2):493–497. doi: 10.1042/bj2330493

Inactivation of 3 alpha-hydroxysteroid dehydrogenase by superoxide radicals. Modification of histidine and cysteine residues causes the conformational change.

H S Kim, P Minard, M D Legoy, D Thomas
PMCID: PMC1153052  PMID: 3006670

Abstract

3 alpha-Hydroxysteroid dehydrogenase (EC 1.1.1.50) from Pseudomonas testosterone was inactivated by superoxide radicals generated by the aerobic xanthine oxidase reaction. Superoxide dismutase, NAD+, bovine serum albumin and histidine and cysteine as free amino acids partially protected the enzyme from inactivation. NADH-binding properties were determined by fluorescence spectroscopy, and no variation was found between native enzyme and the unmodified fraction of the partly inactivated one. The fluorescence emission maximum for the completely inactivated enzyme was shifted 10 nm to a longer wavelength when compared with the native one, and it seems possible that the modification of histidine and cysteine residues by superoxide radicals causes the conformational change of the enzyme and the consequent loss of catalytic activity.

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Selected References

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  1. Blech D. M., Borders C. L., Jr Hydroperoxide anion, HO-2, is an affinity reagent for the inactivation of yeast Cu,Zn superoxide dismutase: modification of one histidine per subunit. Arch Biochem Biophys. 1983 Jul 15;224(2):579–586. doi: 10.1016/0003-9861(83)90245-x. [DOI] [PubMed] [Google Scholar]
  2. Buchanan J. D., Armstrong D. A. The radiolysis of glyceraldehyde-3-phosphate dehydrogenase. Int J Radiat Biol Relat Stud Phys Chem Med. 1978 May;33(5):409–418. doi: 10.1080/09553007814550331. [DOI] [PubMed] [Google Scholar]
  3. Dershwitz M., Novak R. F. Generation of superoxide via the interaction of nitrofurantoin with oxyhemoglobin. J Biol Chem. 1982 Jan 10;257(1):75–79. [PubMed] [Google Scholar]
  4. Freeman B. A., Crapo J. D. Biology of disease: free radicals and tissue injury. Lab Invest. 1982 Nov;47(5):412–426. [PubMed] [Google Scholar]
  5. Fridovich I. Quantitative aspects of the production of superoxide anion radical by milk xanthine oxidase. J Biol Chem. 1970 Aug 25;245(16):4053–4057. [PubMed] [Google Scholar]
  6. Fridovich I. The biology of oxygen radicals. Science. 1978 Sep 8;201(4359):875–880. doi: 10.1126/science.210504. [DOI] [PubMed] [Google Scholar]
  7. Halliwell B., Gutteridge J. M. Oxygen toxicity, oxygen radicals, transition metals and disease. Biochem J. 1984 Apr 1;219(1):1–14. doi: 10.1042/bj2190001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hodgson E. K., Fridovich I. The interaction of bovine erythrocyte superoxide dismutase with hydrogen peroxide: inactivation of the enzyme. Biochemistry. 1975 Dec 2;14(24):5294–5299. doi: 10.1021/bi00695a010. [DOI] [PubMed] [Google Scholar]
  9. Kono Y., Fridovich I. Superoxide radical inhibits catalase. J Biol Chem. 1982 May 25;257(10):5751–5754. [PubMed] [Google Scholar]
  10. Levine R. L., Oliver C. N., Fulks R. M., Stadtman E. R. Turnover of bacterial glutamine synthetase: oxidative inactivation precedes proteolysis. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2120–2124. doi: 10.1073/pnas.78.4.2120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Levine R. L. Oxidative modification of glutamine synthetase. I. Inactivation is due to loss of one histidine residue. J Biol Chem. 1983 Oct 10;258(19):11823–11827. [PubMed] [Google Scholar]
  12. Luedtke R., Karush F. Antibody interaction with a membrane-bound fluorescent ligand on synthetic lipid vesicles. Biochemistry. 1982 Nov 9;21(23):5738–5744. doi: 10.1021/bi00266a002. [DOI] [PubMed] [Google Scholar]
  13. Nishikimi M., Appaji N., Yagi K. The occurrence of superoxide anion in the reaction of reduced phenazine methosulfate and molecular oxygen. Biochem Biophys Res Commun. 1972 Jan 31;46(2):849–854. doi: 10.1016/s0006-291x(72)80218-3. [DOI] [PubMed] [Google Scholar]
  14. STOCKELL A. The binding of diphosphopyridine nucleotide by yeast glyceraldehyde-3-phosphate dehydrogenase. J Biol Chem. 1959 May;234(5):1286–1292. [PubMed] [Google Scholar]
  15. Schneider K., Schlegel H. G. Production of superoxide radicals by soluble hydrogenase from Alcaligenes eutrophus H16. Biochem J. 1981 Jan 1;193(1):99–107. doi: 10.1042/bj1930099. [DOI] [PMC free article] [PubMed] [Google Scholar]

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