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Molecular diversity and double regulatory mechanism of activation of phospholipase C in rat brain.

机译:大鼠脑中磷脂酶C激活的分子多样性和双重调控机制。

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Whereas evidence for a G protein-dependent stimulation of phospholipase C (PLC) is abundant, reports on the inhibition of PLC through a G protein-mediated pathway have only recently begun to appear. In the present study, cerebral cortex membranes were chosen since they have a readily measurable Gpp[NH]p and Ca2+-stimulated PLC activity. Nanomolar concentrations of Gpp[NH]p, a hydrolysis-resistant GTP analogue, inhibited basal inositol 1,4,5-trisphosphate (IP3) production, with a maximum inhibition of 25% at 10 nM. Increasing the concentrations of Gpp[NH]p to over 10 nM resulted in a reversal of the inhibitory effect and onset of stimulation of IP3 production. GDPbetaS as a G protein inhibitor and U-73122 as a putative PLC-beta inhibitor had little effect on basal IP3 production at 100 microM and 1 microM, respectively. However, GDPbetaS and U-73122 completely antagonized both the inhibition and the stimulation of IP3 production produced by lower and higher concentrations, respectively, of Gpp[NH]p. Rat cortical membranes expressed a greater amount of PLC-beta1. These data suggest that PLC-beta1 isozymes may be regulated by both inhibitory and stimulatory G protein-mediated mechanisms.
机译:尽管有大量的依赖G蛋白的磷脂酶C(PLC)刺激的证据,但有关通过G蛋白介导的途径抑制PLC的报道只是最近才开始出现。在本研究中,选择了大脑皮层膜,因为它们具有易于测量的Gpp [NH] p和Ca2 +刺激的PLC活性。耐水解的GTP类似物Gpp [NH] p的纳摩尔浓度可抑制基础肌醇1,4,5-三磷酸(IP3)的生成,在10 nM时最大抑制率为25%。将Gpp [NH] p的浓度增加到10 nM以上会导致抑制作用的逆转和IP3产生刺激的开始。 GDPbetaS作为一种G蛋白抑制剂,U-73122作为一种公认的PLC-beta抑制剂,分别对100 microM和1 microM的基础IP3产生影响很小。但是,GDPbetaS和U-73122完全拮抗了分别由较低和较高浓度的Gpp [NH] p产生的IP3产生的抑制和刺激作用。大鼠皮质膜表达大量的PLC-beta1。这些数据表明,PLC-beta1同工酶可能受抑制和刺激性G蛋白介导的机制。

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