title: Studies on enzyme-substrate interactions of cholinephosphotransferase from rat liver.
abstract: In order to elucidate the reaction mechanism and the substrate-binding sites, CDPcholine:1,2-diacylglycerol cholinephosphotransferase (EC 2.7.8.2), prepared from rat liver microsomal fraction, has been subjected to kinetic analysis and substrate specificity studies. Kinetic evidence supports the hypothesis of a Bi-Bi sequential mechanism, involving a direct nucleophilic attack of diacylglycerol on CDPcholine during the reaction. To investigate the substrate requirements for recognition and catalysis, several CDPcholine analogs, modified in the nitrogen base or in the sugar or in the pyrophosphate bridge, have been synthesized, characterized and assayed as substrates and/or inhibitors of the reaction. The amino group on the pyrimidine ring, the 2'-alcoholic function of the ribose moiety as well as the pyrophosphate bridge have been identified as critical sites for enzyme-substrates interactions.
authors.forename: G
authors.initials: G
authors.lastname: Pontoni
authors.forename: C
authors.initials: C
authors.lastname: Manna
authors.forename: A
authors.initials: A
authors.lastname: Salluzzo
authors.forename: L
authors.initials: L
authors.lastname: del Piano
authors.forename: P
authors.initials: P
authors.lastname: Galletti
authors.forename: M
authors.initials: M
authors.lastname: De Rosa
authors.forename: V
authors.initials: V
authors.lastname: Zappia
mesh_terms: D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002384:Catalysis; D003566:Cytidine Diphosphate Choline / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000378:metabolism; D002798:Diacylglycerol Cholinephosphotransferase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D010770:Phosphotransferases / Q000378:metabolism*; D051381:Rats; D013379:Substrate Specificity
chemical_list: D003566:Cytidine Diphosphate Choline; D010770:Phosphotransferases; D002798:Diacylglycerol Cholinephosphotransferase