Phosphatidylcholine synthesis in castor bean endosperm: the localization and control of CTP: choline-phosphate cytidylyltransferase activity.
The reaction catalyzed by CTP:choline-phosphate cytidylyltransferase (EC 2.7.7.15) has been postulated to be a control reaction in the synthesis of phosphatidylcholine (PtdCho) in many animal tissues and some plants. In 3-day-old castor bean (Ricinus communis L. var. Hale) endosperm the majority of cytidylyltransferase activity resided in a 12,000gav 10-min pellet. Following density-gradient fractionation, 60 to 70% of the enzyme activity was associated with the endoplasmic reticulum (ER) fraction, with the remainder in the particulate fraction being in an unidentified membrane band (band A), less than occurred in the soluble fractions. The properties and kinetics of the forward and reverse reactions are described. About 40% of the total ER activity could be solubilized by treatment of the fraction with 0.32 M KCl, which resulted in a threefold increase in the specific activity of the enzyme. The Michaelis constants of the solubilized enzyme were similar to those of the ER activity. The activity of the solubilized enzyme was stimulated 35% by addition of phosphatidylglycerol or phosphatidylinositol to the assay. Addition of a number of other phospholipids to the incubation medium caused only a small change in activity (+/- 10%) but the enzyme could be stimulated up to 60% by the addition of 0.01-1 mM sodium oleate. A combination of 0.25 mM PtdCho with oleate in the assay resulted in additional stimulation at all concentrations of oleate. Oleate had no effect on the ER activity. These results are discussed in relation to the regulation of cytidylyltransferase activity in plants.
Kinney
A J
AJ
Department of Botany, Louisiana State University, Baton Rouge 70803-1705.
Moore
T S
TS
D002367:Ricinus communis / Q000201:enzymology; D019916:Choline-Phosphate Cytidylyltransferase; D007700:Kinetics; D009713:Nucleotidyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000096:biosynthesis*; D010944:Plants / Q000201:enzymology*; D010947:Plants, Toxic
D010713:Phosphatidylcholines; D009713:Nucleotidyltransferases; D019916:Choline-Phosphate Cytidylyltransferase