Glucose phosphoenolpyruvate-dependent phosphotransferase system of Streptococcus mutans GS5 studied by using cell-free extracts.
The glucose phosphotransferase system (PTS) of Streptococcus mutans GS5 has been partially characterized, using fractions derived from cells treated with the muramidase mutanolysin. Membranes retained functional PTS enzymes for the phosphoenolpyruvate-dependent phosphorylation of glucose, fructose, and mannose. This was confirmed by assaying membranes directly for enzyme I (EI) and enzyme IIglc (EIIglc) by employing specific phosphoryl-exchange reactions for each factor. Membranes prepared from glucose PTS- mutants, however, were either deficient in glucose phosphorylation or reflected the "leakiness" displayed by whole cells. Mutant membranes were unable to catalyze the glucose:glucose 6-phosphate transphosphorylation reaction, indicating a defective EIIglc in these fractions. Although total cellular EI activities in the mutant clones were about the same as that measured for the wild-type strain by employing the pyruvate:phosphoenolpyruvate phosphoryl-exchange reaction, mutant membranes were found to possess less than 10% of the specific EI activity of wild-type membranes. The cytoplasmic fractions of mutants, however, displayed markedly increased specific activities for this enzyme when compared with wild-type extracts. These results strongly suggest a molecular association of EI with a normal membrane protein, perhaps EIIglc, that is absent in mutants. This would explain the absence of fructose PTS activity in glucose PTS- mutant membranes despite the fact that whole cells of these clones are normal for this transport function.
D002250:Carbon Radioisotopes; D002462:Cell Membrane / Q000201:enzymology; D002474:Cell-Free System; D007700:Kinetics; D009154:Mutation; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000378:metabolism*; D013295:Streptococcus mutans / Q000201:enzymology*; D013379:Substrate Specificity
D002250:Carbon Radioisotopes; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C031629:phosphoenolpyruvate-glucose phosphotransferase