Kinetic and biochemical properties of CTP:choline-phosphate cytidylyltransferase from the rat brain.
In order to investigate the mechanisms involved in some brain disorders at the membrane level, we studied the kinetics and biochemical properties of brain CTP:choline-phosphate cytidylyltransferase (EC 2.7.7.15), the rate-limiting enzyme of the two-step biosynthesis of phosphatidylcholine. This enzyme catalyzes the biosynthesis of CDPcholine from choline phosphate and CTP. We found that its subcellular localization (mainly in microsomal and cytosolic fractions) was different from that of phosphatidylethanolamine N-methyltransferase (EC 2.1.1.17), the enzyme of the alternative pathway for phosphatidylcholine synthesis. CTP:choline-phosphate cytidylyltransferase showed a Km of 10 mM for CTP and 0.3 mM for choline phosphate and exhibited a random mechanism. CDPcholine, the reaction product, was a competitive inhibitor of choline phosphate and CTP utilization and had a Ki of 0.090 mM. Both particulate and soluble enzymes required Mg2+ and exhibited an optimal pH at about 7. Cytosolic activity was enhanced by addition of unsaturated fatty acids or phospholipids extracted from brain membranes. Such an enhancement was increased with the centrifugation time used for preparing the soluble enzyme.
Mages
F
F
Unité 205, de l'Institut National de la Santé et de la Recherche Médicale, Institut National des Sciences Appliquées, Villeurbanne, France.
Rey
C
C
Fonlupt
P
P
Pacheco
H
H
D000818:Animals; D001921:Brain / Q000201:enzymology*; D002384:Catalysis; D019916:Choline-Phosphate Cytidylyltransferase; D003566:Cytidine Diphosphate Choline / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D004789:Enzyme Activation / Q000187:drug effects; D007700:Kinetics; D008297:Male; D008780:Methyltransferases / Q000302:isolation & purification; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D010713:Phosphatidylcholines / Q000096:biosynthesis; D010767:Phosphorylcholine / Q000378:metabolism; D051381:Rats; D013347:Subcellular Fractions / Q000201:enzymology
D010713:Phosphatidylcholines; D010767:Phosphorylcholine; D003566:Cytidine Diphosphate Choline; D008780:Methyltransferases; D009713:Nucleotidyltransferases; D019916:Choline-Phosphate Cytidylyltransferase