Kinetic mechanism of choline kinase from rat striata.
The kinetic mechanism of choline kinase associated with both the cytosolic and membrane fractions of synaptosomes isolated from rat striata was studied. The velocity of choline kinase was measured using various concentrations of MgATP at several concentrations of uncomplexed Mg2+ and a single concentration of choline. This experiment was repeated using different concentrations of choline. Analysis of these data according to a terreactant mechanism indicates that MgATP binds in rapid equilibrium prior to Mg2+, but the binding of MgATP and choline is random. Product inhibition by phosphorylcholine was noncompetitive versus both choline and MgATP. Hemicholinium-3 (HC-3), an analog of choline and competitive inhibitor of the sodium-dependent high affinity choline transport system, was noncompetitive versus choline and uncompetitive versus MgATP at high levels of Mg2+. However, when the concentration of Mg2+ was decreased below the KMg2 +, HC-3 was noncompetitive versus MgATP. Thiocholine, another analog of choline, gave slope-linear intercept hyperbolic inhibition versus choline. Mg-5'-adenylyl imidodiphosphate, an analog of MgATP, was competitive versus MgATP and noncompetitive versus choline. Virtually identical results were obtained using either soluble or particulate forms of choline kinase from rat striata. All data are consistent with the mechanism suggested by initial velocity studies alone and additionally suggest that the release of MgADP is slow, occurs last, and may limit the overall rate of the reaction.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002797:Choline Kinase / Q000378:metabolism*; D003342:Corpus Striatum / Q000201:enzymology*; D006426:Hemicholinium 3 / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D008433:Mathematics; D010767:Phosphorylcholine / Q000494:pharmacology; D010770:Phosphotransferases / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013860:Thiocholine / Q000494:pharmacology
D010767:Phosphorylcholine; D006426:Hemicholinium 3; D013860:Thiocholine; D000255:Adenosine Triphosphate; D010770:Phosphotransferases; D002797:Choline Kinase