Analysis of the water-soluble products of phosphatidylcholine breakdown by ion-exchange chromatography. Bombesin and TPA (12-O-tetradecanoylphorbol 13-acetate) stimulate choline generation in Swiss 3T3 cells by a common mechanism.
A method for the rapid and quantitative separation of glycerophosphocholine, choline phosphate and choline upon ion-exchange columns is described. The method has been utilized to examine the stimulation of phosphatidylcholine breakdown in quiescent Swiss 3T3 cells in response to bombesin and 12-O-tetradecanoylphorbol 13-acetate (TPA). The stimulated generation of choline is shown to precede that of choline phosphate, with no effect upon glycerophosphocholine levels; but was attenuated in cells in which protein kinase C activity was down-regulated. The results thus suggest that stimulation of the cells with either bombesin or TPA activates phospholipase D-catalysed phosphatidylcholine breakdown by a common mechanism involving the activation of protein kinase C.
Cook
S J
SJ
Department of Biochemistry, University of Glasgow, Scotland, U.K.
Wakelam
M J
MJ
D001839:Bombesin / Q000494:pharmacology*; D002460:Cell Line; D002794:Choline / Q000302:isolation & purification / Q000378:metabolism; D002852:Chromatography, Ion Exchange; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D005997:Glycerylphosphorylcholine / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D010713:Phosphatidylcholines / Q000378:metabolism*; D010767:Phosphorylcholine / Q000302:isolation & purification / Q000378:metabolism; D012995:Solubility; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014316:Tritium; D014867:Water
D010713:Phosphatidylcholines; D014867:Water; D014316:Tritium; D010767:Phosphorylcholine; D005997:Glycerylphosphorylcholine; D002794:Choline; D013755:Tetradecanoylphorbol Acetate; D001839:Bombesin