The stimulation and binding of CTP: phosphorylcholine cytidylyltransferase by phosphatidylcholine-oleic acid vesicles.
The activity of the low molecular weight form of cytidylyltransferase from fetal lung cytosol and adult liver cytosol was stimulated more by phosphatidylcholine-oleic acid (1:1 molar ratio) vesicles than by phosphatidylglycerol vesicles. Phosphatidylcholine alone did not stimulate the activity, while oleic acid alone produced only slight stimulation. Vesicles prepared from phosphatidylinositol, phosphatidylglycerol-cholesterol (2:1) and phosphatidylglycerol-phosphatidylcholine (1:1) all stimulated the activity to the same extent. Phosphatidylcholine-oleic acid vesicles (molar ratio 2:1) produced less stimulation than 1:1 vesicles. Phosphatidylcholine-palmitic acid vesicles (2:1) were about 50% as active as the corresponding phosphatidylcholine-oleic acid vesicles. All vesicles were in the size range of small unilamellar vesicles as judged by Sephacryl S-1000 chromatography. Stimulation also occurred when phosphatidylcholine vesicles and oleic acid were added separately to the assay. The stimulation by phospholipid vesicles was correlated with the ability of the vesicles to bind cytidylyltransferase, determined by sucrose density centrifugation of the enzyme-vesicles mixtures. We conclude that the stimulation of soluble cytidylyltransferase occurs through binding of the enzyme to anionic membrane surfaces. Suitable anionic membranes can be prepared either from anionic phospholipids, or by the addition of anionic lipids (unesterified fatty acids or phosphatidylglycerol) to phosphatidylcholine.
Feldman
D A
DA
Rounsifer
M E
ME
Weinhold
P A
PA
D000818:Animals; D002499:Centrifugation, Density Gradient; D019916:Choline-Phosphate Cytidylyltransferase; D002850:Chromatography, Gel; D003570:Cytidine Triphosphate / Q000378:metabolism; D005260:Female; D007700:Kinetics; D008563:Membrane Lipids / Q000378:metabolism*; D009713:Nucleotidyltransferases / Q000378:metabolism*; D019301:Oleic Acid; D009829:Oleic Acids / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000378:metabolism*; D051381:Rats
D008563:Membrane Lipids; D009829:Oleic Acids; D010713:Phosphatidylcholines; D019301:Oleic Acid; D003570:Cytidine Triphosphate; D009713:Nucleotidyltransferases; D019916:Choline-Phosphate Cytidylyltransferase