Studies on CDP-choline:1,2-diacylglycerol cholinephosphotransferase activity in rat arterial wall.
The properties of CDP-choline:1,2-diacylglycerol cholinephosphotransferase (CPT) (EC 2.7.8.2.), which catalyzes de novo synthesis of phosphatidylcholine, were studied in rat arterial wall. The optimal pH of CPT of the arterial wall was about 8.5. On subcellular fractionation of the arterial wall, the highest activity was found in the microsome-rich fraction; the cytosolic fraction showed only a trace of activity. The Michaelis constant (KM) for CDP-choline was 0.019 mM. The CPT activity of a homogenate of arterial wall increased linearly with increase in concentration of diolein up to 3.2 mM. 20 mM magnesium and 0.2 mM manganese ions caused marked activation respectively and essential for the activity. Calcium, barium, cobalt, copper, and ferrous ions were inhibitory. 0.5 mM ethylenediaminetetraacetic acid (EDTA) and 0.5 mM glycoletherdiamine-N,N,N'N'-tetraacetic acid (GEDTA) increased the activity in the presence of 10 mM magnesium ion. Sonication of the enzyme solution and addition of high concentration of detergent, such as Triton X-100 and Tween 20, markedly decreased the activity. Porcine liver phosphatidylcholine, phosphatidylethanolamine, and especially polyenephosphatidylcholine increased CPT activity of the arterial wall, while lysophosphatidylcholine was strongly inhibitory. The properties of arterial CPT activity under various conditions are discussed.
D000818:Animals; D001011:Aorta / Q000378:metabolism*; D001464:Barium / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D002794:Choline / Q000031:analogs & derivatives*; D003566:Cytidine Diphosphate Choline / Q000378:metabolism*; D002798:Diacylglycerol Cholinephosphotransferase / Q000378:metabolism*; D004075:Diglycerides / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000494:pharmacology; D008297:Male; D008345:Manganese / Q000494:pharmacology; D017830:Octoxynol; D010713:Phosphatidylcholines / Q000096:biosynthesis; D010770:Phosphotransferases / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains
D004075:Diglycerides; D010713:Phosphatidylcholines; D001464:Barium; D011092:Polyethylene Glycols; D008345:Manganese; D003566:Cytidine Diphosphate Choline; D017830:Octoxynol; D004492:Edetic Acid; D010770:Phosphotransferases; D002798:Diacylglycerol Cholinephosphotransferase; D008274:Magnesium; D002794:Choline; D002118:Calcium; C013965:diolein