Oxidases and reductases are involved in metronidazole sensitivity in Helicobacter pylori.
Helicobacter pylori is a contributing factor to the development of gastric and duodenal ulcers and some gastric cancers. Some therapeutic regimes comprise of a number of components, one of which is the antimicrobial metronidazole. A problem with these therapies is the increasing prevalence of metronidazole-resistant (MtrR) H. pylori strains. Several resistance mechanisms have been proposed, and this study addresses the 'scavenging of oxygen' hypothesis. Spectrophotometric assays of cytosolic fractions indicated that metronidazole-sensitive (MtrS) H. pylori isolates had 2.6-fold greater nicotinamide adenine dinucleotide (NADH) oxidase activity, 34-fold greater NADH nitroreductase activity, and eightfold greater nicotinamide adenine dinucleotide phosphate (NADPH) nitroreductase activity than cytosolic fractions from matched MtrR strains. Electrophoresis of cytosolic fractions in non-denaturing gels showed up to 10 protein bands when stained with Coomassie blue. Activity staining of non-denaturing, non-reducing polyacrylamide gels detected NAD(P)H oxidase, disulphide reductase, tetrazolium reductase and nitroreductase activities in the protein bands. Oxidase and reductase activities observed in a band from MtrS strains were absent in the corresponding band from MtrR strains. This band comprised at least 13 proteins, and the major constituent was identified as an alkyl hydroperoxide reductase AhpC subunit. The absence of oxidase and reductase activities in the band from MtrR strains indicated a correlation between the activity of the proteins in this band and the metronidazole-sensitive phenotype.
Trend
M A
MA
School of Biochemistry and Molecular Genetics, The University of New South Wales, NSW 2052, Sydney, Australia.
Jorgensen
M A
MA
Hazell
S L
SL
Mendz
G L
GL
D000595:Amino Acid Sequence; D003600:Cytosol / Q000201:enzymology; D004352:Drug Resistance, Microbial; D004591:Electrophoresis, Polyacrylamide Gel; D016480:Helicobacter pylori / Q000187:drug effects*; D006801:Humans; D008795:Metronidazole / Q000494:pharmacology*; D009243:NAD / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000502:physiology*; D019255:NADPH Oxidases; D010544:Peroxidases / Q000378:metabolism; D054464:Peroxiredoxins; D016415:Sequence Alignment
D009243:NAD; D008795:Metronidazole; D010544:Peroxidases; D054464:Peroxiredoxins; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases