The bacterial phosphotransferase system: Kinetic characterization of the glucose, mannitol, glucitol, and N‐acetylglucosamine systems

Abstract

The kinetic mechanisms by which the glucose, glucitol, N-acetylglucosamine, and mannitol enzymes II catalyze sugar phosphorylation have been investigated in vitro. Lineweaver-Burk analyses indicate that the glucose and glucitol enzymes II catalyze sugar phosphorylation by a sequential mechanism when the two substrates are phospho-enzyme III and sugar. The N-acetylglucosamine and mannitol enzymes II, which do not function with an enzyme III, catalyze sugar phosphorylation by a ping-pong mechanism when the two substrates are phospho-HPr and sugar. These results, as well as previously published kinetic characterizations, suggest a common kinetic mechanism for all enzymes II of the system. It is suggested that all enzymes II and enzyme II-III pairs arose from a single (fused) gene product containing two sites of phosphorylation and that phosphoryl transfer from the second phosphorylation site to sugar can only occur when the enzyme II-III pair is present in the associated state.