Abstract
Anaerobically induced NAD-linked glycerol dehydrogenase of Klebsiella pneumoniae for fermentative glycerol utilization was reported previously to be inactivated in the cell during oxidative metabolism. In vitro inactivation was observed in this study by incubating the purified enzyme in the presence of O2, Fe2+, and ascorbate or dihydroxyfumarate. It appears that O2 and the reducing agent formed H2O2 and that H2O2 reacted with Fe2+ to generate an activated species of oxygen which attacked the enzyme. The in vitro-oxidized enzyme, like the in vivo-inactivated enzyme, showed an increased Km for NAD (but not glycerol) and could no longer be activated by Mn2+ which increased the Vmax of the native enzyme but decreased its apparent affinity for NAD. Ethanol dehydrogenase and 1,3-propanediol oxidoreductase, two enzymes with anaerobic function, also lost activity when the cells were incubated aerobically with glucose. However, glucose 6-phosphate dehydrogenase (NADP-linked), isocitrate dehydrogenase, and malate dehydrogenase, expected to function both aerobically and anaerobically, were not inactivated. Thus, oxidative modification of proteins in vivo might provide a mechanism for regulating the activities of some anaerobic enzymes.
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Selected References
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- Asada K., Yoshikawa K., Takahashi M., Maeda Y., Enmanji K. Superoxide dismutases from a blue-green alga, Plectonema boryanum. J Biol Chem. 1975 Apr 25;250(8):2801–2807. [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Forage R. G., Foster M. A. Glycerol fermentation in Klebsiella pneumoniae: functions of the coenzyme B12-dependent glycerol and diol dehydratases. J Bacteriol. 1982 Feb;149(2):413–419. doi: 10.1128/jb.149.2.413-419.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Forage R. G., Lin E. C. DHA system mediating aerobic and anaerobic dissimilation of glycerol in Klebsiella pneumoniae NCIB 418. J Bacteriol. 1982 Aug;151(2):591–599. doi: 10.1128/jb.151.2.591-599.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fridovich I. The biology of oxygen radicals. Science. 1978 Sep 8;201(4359):875–880. doi: 10.1126/science.210504. [DOI] [PubMed] [Google Scholar]
- Fucci L., Oliver C. N., Coon M. J., Stadtman E. R. Inactivation of key metabolic enzymes by mixed-function oxidation reactions: possible implication in protein turnover and ageing. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1521–1525. doi: 10.1073/pnas.80.6.1521. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gregory E. M., Dapper C. H. Isolation of iron-containing superoxide dismutase from Bacteroides fragilis: reconstitution as a Mn-containing enzyme. Arch Biochem Biophys. 1983 Jan;220(1):293–300. doi: 10.1016/0003-9861(83)90413-7. [DOI] [PubMed] [Google Scholar]
- Gregory E. M., Fridovich I. Induction of superoxide dismutase by molecular oxygen. J Bacteriol. 1973 May;114(2):543–548. doi: 10.1128/jb.114.2.543-548.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gregory E. M., Yost F. J., Jr, Fridovich I. Superoxide dismutases of Escherichia coli: intracellular localization and functions. J Bacteriol. 1973 Sep;115(3):987–991. doi: 10.1128/jb.115.3.987-991.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hassan H. M., Fridovich I. Regulation of the synthesis of superoxide dismutase in Escherichia coli. Induction by methyl viologen. J Biol Chem. 1977 Nov 10;252(21):7667–7672. [PubMed] [Google Scholar]
- Johnson E. A., Burke S. K., Forage R. G., Lin E. C. Purification and properties of dihydroxyacetone kinase from Klebsiella pneumoniae. J Bacteriol. 1984 Oct;160(1):55–60. doi: 10.1128/jb.160.1.55-60.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kanematsu S., Asada K. Ferric and manganic superoxide dismutases in Euglena gracilis. Arch Biochem Biophys. 1979 Jul;195(2):535–545. doi: 10.1016/0003-9861(79)90380-1. [DOI] [PubMed] [Google Scholar]
- LIN E. C., LEVIN A. P., MAGASANIK B. The effect of aerobic metabolism on the inducible glycerol dehydrogenase of Aerobacter aerogenes. J Biol Chem. 1960 Jun;235:1824–1829. [PubMed] [Google Scholar]
- LIN E. C., MAGASANIK B. The activation of glycerol dehydrogenase from Aerobacter aerogenes by monovalent cations. J Biol Chem. 1960 Jun;235:1820–1823. [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Levine R. L. Mixed-function oxidation of histidine residues. Methods Enzymol. 1984;107:370–376. doi: 10.1016/0076-6879(84)07025-7. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Lin E. C. Glycerol dissimilation and its regulation in bacteria. Annu Rev Microbiol. 1976;30:535–578. doi: 10.1146/annurev.mi.30.100176.002535. [DOI] [PubMed] [Google Scholar]
- McGregor W. G., Phillips J., Suelter C. H. Purification and kinetic characterization of a monovalent cation-activated glycerol dehydrogenase from Aerobacter aerogenes. J Biol Chem. 1974 May 25;249(10):3132–3139. [PubMed] [Google Scholar]
- McPhedran P., Sommer B., Lin E. C. CONTROL OF ETHANOL DEHYDROGENASE LEVELS IN AEROBACTER AEROGENES. J Bacteriol. 1961 Jun;81(6):852–857. doi: 10.1128/jb.81.6.852-857.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Milne J. A., Cook R. A. Role of metal cofactors in enzyme regulation. Differences in the regulatory properties of the Escherichia coli nicotinamide adenine dinucleotide specific malic enzyme depending on whether Mg2+ or Mn2+ serves as divalent cation. Biochemistry. 1979 Aug 7;18(16):3604–3610. doi: 10.1021/bi00583a026. [DOI] [PubMed] [Google Scholar]
- Nettleton C. J., Bull C., Baldwin T. O., Fee J. A. Isolation of the Escherichia coli iron superoxide dismutase gene: evidence that intracellular superoxide concentration does not regulate oxygen-dependent synthesis of the manganese superoxide dismutase. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4970–4973. doi: 10.1073/pnas.81.15.4970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Orsi B. A., Cleland W. W. Inhibition and kinetic mechanism of rabbit muscle glyceraldehyde-3-phosphate dehydrogenase. Biochemistry. 1972 Jan 4;11(1):102–109. doi: 10.1021/bi00751a018. [DOI] [PubMed] [Google Scholar]
- Que L., Jr, Widom J., Crawford R. L. 3,4-Dihydroxyphenylacetate 2,3-dioxygenase. A manganese(II) dioxygenase from Bacillus brevis. J Biol Chem. 1981 Nov 10;256(21):10941–10944. [PubMed] [Google Scholar]
- Ruch F. E., Jr, Lin E. C., Kowit J. D., Tang C. T., Goldberg A. L. In vivo inactivation of glycerol dehydrogenase in Klebsiella aerogenes: properties of active and inactivated proteins. J Bacteriol. 1980 Mar;141(3):1077–1085. doi: 10.1128/jb.141.3.1077-1085.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ruch F. E., Lengeler J., Lin E. C. Regulation of glycerol catabolism in Klebsiella aerogenes. J Bacteriol. 1974 Jul;119(1):50–56. doi: 10.1128/jb.119.1.50-56.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ruch F. E., Lin E. C. Independent constitutive expression of the aerobic and anaerobic pathways of glycerol catabolism in Klebsiella aerogenes. J Bacteriol. 1975 Oct;124(1):348–352. doi: 10.1128/jb.124.1.348-352.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Silver S., Johnseine P., King K. Manganese Active Transport in Escherichia coli. J Bacteriol. 1970 Dec;104(3):1299–1306. doi: 10.1128/jb.104.3.1299-1306.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sridhara S., Wu T. T., Chused T. M., Lin E. C. Ferrous-activated nicotinamide adenine dinucleotide-linked dehydrogenase from a mutant of Escherichia coli capable of growth on 1, 2-propanediol. J Bacteriol. 1969 Apr;98(1):87–95. doi: 10.1128/jb.98.1.87-95.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tanaka S., Lerner S. A., Lin E. C. Replacement of a phosphoenolpyruvate-dependent phosphotransferase by a nicotinamide adenine dinucleotide-linked dehydrogenase for the utilization of mannitol. J Bacteriol. 1967 Feb;93(2):642–648. doi: 10.1128/jb.93.2.642-648.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]