Direct evidence for phorbol ester-stimulated accumulation of diacylglycerol derived from phosphatidylcholine.
A fluorescent analog of phosphatidylcholine, 1-acyl-2-[N-(4-nitrobenzo-2-oxa-1,3-diazole)aminocaproyl]phosphatidyl choline (NBD-PC), was inserted into the plasma membrane of HeLa cells. Treatment of NBD-PC-labelled cells with the tumour promoting phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), resulted in the production of fluorescent diacylglycerol, 1-acyl-2-[N-(4-nitrobenzo-2-oxa-1,3-diazole)-aminocaproyl]diacylglycerol (NBD-DG), suggesting that phorbol ester activates a PC-specific phospholipase C in these cells. Furthermore pretreatment of cells with phorbol-12,13-dibutyrate for 24 h, a process which depletes cellular protein kinase C activity, inhibited the ability of TPA to induce this effect. These results suggest that TPA stimulation of a phospholipase C specific for PC in HeLa cells is mediated via protein kinase C.
Glatz
J A
JA
School of Biological Sciences, Flinders University of South Australia, Bedford Park.
Muir
J G
JG
Murray
A W
AW
D002462:Cell Membrane / Q000378:metabolism; D002855:Chromatography, Thin Layer; D004075:Diglycerides / Q000378:metabolism*; D005989:Glycerides / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D010703:Phorbol Esters / Q000494:pharmacology*; D010713:Phosphatidylcholines / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D004075:Diglycerides; D005989:Glycerides; D010703:Phorbol Esters; D010713:Phosphatidylcholines; D013755:Tetradecanoylphorbol Acetate