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. 1985 Jul;163(1):243–247. doi: 10.1128/jb.163.1.243-247.1985

Evidence for covalently cross-linked dimers and trimers of enzyme I of the Escherichia coli phosphotransferase system.

F C Grenier, J Reizer, E B Waygood, M H Saier Jr
PMCID: PMC219104  PMID: 3891729

Abstract

Enzyme I of the bacterial phosphotransferase system catalyzes transfer of the phosphoryl moiety from phosphoenolpyruvate to both of the heat-stable phosphoryl carrier proteins of the phosphotransferase system, HPr and FPr. Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and high-pressure liquid chromatography, we demonstrated the existence of covalently cross-linked enzyme I dimers and trimers. Enzyme I exchange assays and phosphorylation experiments with [32P]phosphoenolpyruvate showed that covalent dimers and trimers are catalytically active. Inhibitors of the enzyme I-catalyzed phosphoenolpyruvate-pyruvate exchange block the phosphorylation of enzyme I dimers and trimers. Inhibition of the activity of enzyme I by N-ethylmaleimide, but not that by p-chloromercuriphenylsulfonate, could be overcome by high concentrations of enzyme, suggesting that N-ethylmaleimide modification changes the associative properties of enzyme I. We present evidence for two distinct classes of sulfhydryl groups in enzyme I.

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Selected References

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

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