title: The phosphoenolpyruvate-dependent fructose-specific phosphotransferase system in Rhodopseudomonas sphaeroides. Mechanism for transfer of the phosphoryl group from phosphoenolpyruvate to fructose.
abstract: The phosphoenolpyruvate-dependent fructose-specific phosphotransferase system in Rhodopseudomonas sphaeroides is a membrane-bound complex of two enzymes, an integral membrane protein EII and a soluble factor SF. In media of high ionic strength the binding constant of SF to the membranes is 55 nM. Phosphorylation of SF, the first step in the reaction sequence, has no influence on the binding. The second step is the transfer of the phosphoryl group from SF to EII. The physical existence of both phosphorylated SF and EII is demonstrated and it is shown that the phosphoryl group is donated to the next species in the reaction sequence. The molecular mass of SF is 110 kDa. EII is almost completely extracted from the membrane as a dimer. The molecular mass of the monomer is 55 kDa. Both SF and EII possess thiols that are essential for catalysis. The thiol on EII is protected against inactivation by N-ethylmaleimide in the phosphorylated state. Kinetic experiments show that the binding site for fructose on EII is induced by phosphorylation of EII ('ping-pong' mechanism). The affinity constants of the phosphotransferase complex for phosphoenolpyruvate and fructose at infinite concentration of the other substrate are 25 microM and 8 microM, respectively. The fructose phosphorylation rate equation is given as a function of the concentration of the two enzymes and the two substrates.
mesh_terms: D001345:Autoradiography; D002384:Catalysis; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008956:Models, Chemical; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000378:metabolism*; D010766:Phosphorylation; D012242:Rhodobacter sphaeroides / Q000201:enzymology* / Q000254:growth & development; D012995:Solubility; D013379:Substrate Specificity
chemical_list: D010731:Phosphoenolpyruvate Sugar Phosphotransferase System