Bacterial phosphotransferase system. Solubilization and purification of the glucose-specific enzyme II from membranes of Salmonella typhimurium.
The glucose-specific enzyme II (IIGlc) of the phosphoenolpyruvate-dependent phosphotransferase system of Salmonella typhimurium has been purified to homogeneity. Purification included the following steps: detergent solubilization of membranes in polydisperse octyloligooxyethylene, isoelectrofocusing, chromatofocusing, and either glycerol gradient centrifugation or gel filtration, all in the presence of the same detergent. Enzymatic activity was assayed as phosphoenolpyruvate-dependent phosphorylation of methyl-alpha-D-glucopyranoside. It could be measured after detergent dilution only and required the presence of phosphatidylglycerol in a sonicated suspension. An antiserum prepared against enzyme IIGlc specifically inhibited phosphorylation of methyl-alpha-D-glucopyranoside. In the solubilized state, purified enzyme IIGlc exists as a complex of molecular weight of 105,000 and a sedimentation coefficient of 3.8 S. In polyacrylamide gels in sodium dodecyl sulfate, it has an apparent molecular weight of about 40,000.
D000596:Amino Acids / Q000378:metabolism; D000936:Antigen-Antibody Complex; D002462:Cell Membrane / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D007106:Immune Sera; D007700:Kinetics; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000302:isolation & purification* / Q000378:metabolism; D012486:Salmonella typhimurium / Q000201:enzymology*
D000596:Amino Acids; D000936:Antigen-Antibody Complex; D007106:Immune Sera; D010446:Peptide Fragments; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C031629:phosphoenolpyruvate-glucose phosphotransferase