Cook S J, Wakelam M J
Department of Biochemistry, University of Glasgow, Scotland, U.K.
Biochem J. 1989 Oct 15;263(2):581-7. doi: 10.1042/bj2630581.
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.
本文描述了一种在离子交换柱上快速定量分离甘油磷酸胆碱、磷酸胆碱和胆碱的方法。该方法已被用于检测在静止的瑞士3T3细胞中,蛙皮素和12-O-十四酰佛波醇-13-乙酸酯(TPA)对磷脂酰胆碱分解的刺激作用。结果表明,胆碱的刺激生成先于磷酸胆碱,对甘油磷酸胆碱水平无影响;但在蛋白激酶C活性下调的细胞中,这种刺激作用减弱。因此,结果表明用蛙皮素或TPA刺激细胞,通过涉及蛋白激酶C激活的共同机制,激活磷脂酶D催化的磷脂酰胆碱分解。