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Identification of an integral plasma membrane pool of protein kinase C in mammalian tissues and cells

机译:鉴定哺乳动物组织和细胞中蛋白激酶C完整的质膜池

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摘要

Protein kinase C (PKC) is a family of serine/threonine protein kinases that are pivotal in cellular regulation. Since its discovery in 1977, PKCs have been known as cytosolic and peripheral membrane proteins. However, there are reports that PKC can insert into phospholipids vesicles in vitro. Given the intimate relationship between the plasma membrane and the activation of PKC, it is important to determine whether such "membrane-inserted" form of PKC exists in mammalian cells or tissues. Here, we report the identification of an integral plasma membrane pool for all the 10 PKC isozymes in vivo by their ability to partition into the detergent-rich phase in Triton X-114 phase partitioning, and by their resistance to extractions with 0.2 M sodium carbonate (pH 11.5), 2 M urea and 2 M sodium chloride. The endogenous integral membrane pool of PKC in mouse fibroblasts is found to be acutely regulated by phorbol ester or diacylglycerol, suggesting that this pool of PKC may participate in cellular processes known to be regulated by PKC. At least for PKC alpha, the C2-V3 region at the regulatory domain of the kinase is responsible for membrane integration. Further exploration of the function of this novel integral plasma membrane pool of PKC will not only shed new light on molecular mechanisms underlying its cellular functions but also provide new strategies for pharmaceutical modulation of this important group of kinases. (c) 2004 Elsevier Inc. All rights reserved.
机译:蛋白激酶C(PKC)是丝氨酸/苏氨酸蛋白激酶家族,在细胞调节中起关键作用。自1977年发现以来,PKC被称为胞质和外周膜蛋白。但是,有报道说PKC可以在体外插入磷脂囊泡中。考虑到质膜和PKC活化之间的密切关系,重要的是要确定这种“膜插入”形式的PKC是否存在于哺乳动物细胞或组织中。在这里,我们报告了通过在Triton X-114相分配中分配入富含去污剂的相的能力,以及它们对用0.2 M碳酸钠萃取的抗性,鉴定了体内所有10种PKC同工酶的完整质膜池(pH 11.5),2 M尿素和2 M氯化钠。发现小鼠成纤维细胞中PKC的内源性整体膜池受佛波酯或二酰基甘油的急性调控,这表明该PKC池可能参与已知受PKC调控的细胞过程。至少对于PKCα,激酶调节域的C2-V3区负责膜整合。对这种新型的整体PKC质膜池功能的进一步探索,不仅将为揭示其细胞功能的分子机制提供新的思路,而且还将为这一重要的激酶组的药物调节提供新的策略。 (c)2004 Elsevier Inc.保留所有权利。

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