Bacterial phosphotransferase system: regulation of the glucose and mannose enzymes II by sulfhydryl oxidation.
We have investigated the effect of oxidizing agents on methyl alpha-glucoside phosphorylation by the Escherichia coli phosphotransferase system (PTS). Oxidizing agents inhibited methyl alpha-glucoside phosphorylation at low methyl alpha-glucoside concentrations, and the degree of inhibition was shown to decrease with increasing concentrations of methyl alpha-glucoside. Results of studies with mutant bacteria and substrate analogues of the glucose and mannose enzymes II showed that contrary to the interpretation of Robillard and Konings [Robillard, G. T., & Konings, W. N. (1981) Biochemistry 20, 5025-5032] the apparent change in the Km value for methyl alpha-glucoside phosphorylation induced by sulfhydryl oxidation is not due to the formation of a low-affinity, oxidized form of the glucose enzyme II. Rather, the results are explained by the presence of two phosphotransferase systems that phosphorylate methyl alpha-glucoside with different affinities and that are differentially sensitive to oxidizing agents. The low Km system corresponds to the glucose enzyme II, which is strongly inhibited by potassium ferricyanide, phenazine methosulfate, and plumbagin. The high Km system corresponds to the mannose enzyme II, which is less sensitive to inhibition by these oxidizing agents. This differential sensitivity to inhibition by oxidizing agents can account for the apparent Km change for methyl alpha-glucoside phosphorylation reported by Robillard and Konings. The physiological significance of sulfhydryl oxidation in the enzymes II of the PTS has yet to be ascertained.
D003847:Deoxyglucose / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D005292:Ferricyanides / Q000494:pharmacology; D005295:Ferrocyanides / Q000494:pharmacology; D007700:Kinetics; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000378:metabolism*; D010766:Phosphorylation; D012486:Salmonella typhimurium / Q000201:enzymology*
D005292:Ferricyanides; D005295:Ferrocyanides; C007931:hexacyanoferrate III; D003847:Deoxyglucose; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C031629:phosphoenolpyruvate-glucose phosphotransferase; C037992:phosphoenolpyruvate-mannose phosphotransferase; C020354:hexacyanoferrate II