The enhancement of phosphatidylcholine biosynthesis by angiotensin II in H9c2 cells.
The effect of angiotensin II on the biosynthesis of phosphatidylcholine in rat heart myoblastic (H9c2) cells was investigated. Cells were incubated with [methyl-3H]choline, and the labelling of phosphatidylcholine at different time intervals was examined. When cells were pretreated with angiotensin II, a significant increase in the labelling of phosphatidylcholine was observed. Analysis of the labelled phosphatidylcholine precursors indicated that the conversion of phosphocholine to CDP-choline was enhanced by angiotensin II treatment. Determination of enzyme activities in the CDP-choline pathway revealed that the activities of choline kinase or CDP-choline: diacylglycerol cholinephosphotransferase were not changed, but the activities of CTP:phosphocholine cytidylyltransferase were stimulated in both the particulate and soluble fractions. The stimulation of the cytidylyltransferase by angiotensin II was not abolished by okadaic acid, indicating that the activation of the enzyme was not mediated via the okadaic-sensitive dephosphorylation mechanism. Alternatively, the stimulation of the cytidylyltransferase activity was completely abolished by protein kinase C inhibitors. Immunoblotting studies revealed that levels of the cytidylyltransferase in the soluble and particulate fractions were not affected by angiotensin II treatment. We conclude that the increase in phosphatidylcholine biosynthesis by angiotensin II was a direct result of the enhancement of the cytidylyltransferase activity. The enhancement of enzyme activity was not mediated via enzyme translocation, but by a mechanism which was intimately associated with the protein kinase C cascade.
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Department of Biochemistry and Molecular Biology, University of Manitoba, Winnipeg, Canada.
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D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D002460:Cell Line; D002794:Choline / Q000378:metabolism; D019916:Choline-Phosphate Cytidylyltransferase; D003566:Cytidine Diphosphate Choline / Q000378:metabolism; D002798:Diacylglycerol Cholinephosphotransferase / Q000378:metabolism; D004075:Diglycerides / Q000032:analysis / Q000378:metabolism; D004988:Ethers, Cyclic / Q000494:pharmacology; D009206:Myocardium / Q000378:metabolism*; D009713:Nucleotidyltransferases / Q000378:metabolism; D019319:Okadaic Acid; D010713:Phosphatidylcholines / Q000096:biosynthesis*; D047428:Protein Kinase Inhibitors; D051381:Rats
D004075:Diglycerides; D004988:Ethers, Cyclic; D010713:Phosphatidylcholines; D047428:Protein Kinase Inhibitors; D000804:Angiotensin II; D019319:Okadaic Acid; D003566:Cytidine Diphosphate Choline; D009713:Nucleotidyltransferases; D019916:Choline-Phosphate Cytidylyltransferase; D002798:Diacylglycerol Cholinephosphotransferase; D002794:Choline