Hydrolysis of cell surface inositol phospholipid leads to the delayed stimulation of phosphatidylinositol synthesis in bovine aortic endothelial cells.
In order to address the issue of how inositol phospholipid synthesis is controlled in a resting cell we looked for enhanced [3H]phosphatidylinositol (PtdIns) labelling in response to the hydrolysis of cell surface PtdIns. Bacillus thuringiensis PtdIns-PLC when added to intact bovine aortic endothelial (BAE) cells rapidly hydrolysed 9.1 +/- 1% of the total cellular PtdIns. This result suggests that BAE cells have a cell surface pool of PTdIns. Hydrolysis of cell surface PtdIns, in contrast to the agonist-stimulated hydrolysis of inner leaflet PtdIns, did not lead to a rapid (minutes) stimulation of PtdIns resynthesis. Prolonged incubation of BAE cells with PtdIns-PLC led to further hydrolysis of PtdIns (up to 20% of total cellular PtdIns). This second phase of PtdIns-PLC induced hydrolysis was inhibited by the addition of brefeldin A suggesting that it was dependent on vesicular traffic to the plasma membrane from the endoplasmic reticulum. Furthermore, the above result suggests that prolonged incubation of intact cells with PtdIns-PLC leads to the slow depeletion of intracellular PtdIns stores. This second phase of PtdIns-PLC induced hydrolysis was associated with PtdIns resynthesis since prolonged incubation with PtdIns-PLC, but not B. cereus PtdCho-PLC (which does not hydrolyse PtdIns), led to enhanced PtdIns labelling. The results indicate that extracellular PtdIns-PLC induced PtdIns resynthesis may occur due to PtdIns-PLC induced intracellular PtdIns depletion.
Sillence
D J
DJ
Department of Physiology and Cellular Biophysics, College of Physicians and Surgeons of Columbia University, New York, NY 10032.
Low
M G
MG
D000818:Animals; D020126:Brefeldin A; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D003517:Cyclopentanes / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000378:metabolism*; D006868:Hydrolysis / Q000187:drug effects; D010713:Phosphatidylcholines / Q000494:pharmacology; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D010716:Phosphatidylinositols / Q000096:biosynthesis* / Q000494:pharmacology; D010727:Phosphoric Diester Hydrolases / Q000494:pharmacology; D011433:Propranolol / Q000494:pharmacology; D010738:Type C Phospholipases / Q000378:metabolism / Q000494:pharmacology
D003517:Cyclopentanes; D010713:Phosphatidylcholines; D010716:Phosphatidylinositols; D020126:Brefeldin A; D011433:Propranolol; D010727:Phosphoric Diester Hydrolases; D010738:Type C Phospholipases; D043265:Phosphatidylinositol Diacylglycerol-Lyase