Analysis of CaM-kinase signaling in cells.

Analysis of CaM-kinase signaling in cells.
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DOI:
10.1016/j.ceca.2011.02.007
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发表时间:
2011-07
期刊:
影响因子:
4
通讯作者:
Soderling TR
Soderling TR
中科院分区:
生物学2区
文献类型:
--
作者:
Wayman GA;Tokumitsu H;Davare MA;Soderling TR

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细胞内游离钙的变化是一种常见的信号机制,它调节了大多数细胞的一系列生理过程。对细胞内Ca2+增加的反应通常由无处不在的钙调蛋白(CaM)介导,钙调蛋白与Ca2+结合后可以与许多蛋白质相互作用并改变其功能,包括被称为CaM激酶(CaMKs)的蛋白激酶家族。特别令人感兴趣的是多功能camk,如CaMKI、CaMKII、CaMKIV和CaMKK,它们可以磷酸化多个下游靶点。这篇综述将概述我们用来确定哪些成员和/或CaMK家族的亚型介导特异性细胞反应的几种方案,重点是在神经元中的研究。许多先前的研究都依赖于单一的方法,如药理学抑制剂或转染的显性阴性激酶结构。由于这些协议中的每一个都有其局限性,这将被讨论,我们强调在绘制细胞信号通路时使用多种独立方法的必要性。
A change in intracellular free calcium is a common signaling mechanism that modulates a wide array of physiological processes in most cells. Responses to increased intracellular Ca2+ are often mediated by the ubiquitous protein calmodulin (CaM) that upon binding Ca2+ can interact with and alter the functionality of numerous proteins including a family of protein kinases referred to as CaM-kinases (CaMKs). Of particular interest are multifunctional CaMKs, such as CaMKI, CaMKII, CaMKIV and CaMKK, that can phosphorylate multiple downstream targets. This review will outline several protocols we have used to identify which members and/or isoforms of this CaMK family mediate specific cellular responses with a focus on studies in neurons. Many previous studies have relied on a single approach such as pharmacological inhibitors or transfected dominant-negative kinase constructs. Since each of these protocols has its limitations, that will be discussed, we emphasize the necessity to use multiple, independent approaches in mapping out cellular signaling pathways.
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