Reversible translocation of p115-RhoGEF by G12/13-coupled receptors
Reversible translocation of p115-RhoGEF by G12/13-coupled receptors
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DOI：
10.1002/jcb.21732
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发表时间：
2008-08-01
影响因子：
4
通讯作者：
Siehler, Sandra
中科院分区：
文献类型：
--
作者：
Meyer, Bruno H.;Freuler, Felix;Siehler, Sandra
G protein-coupled receptors (GPCRs) are important targets for medicinal agents.四种不同的 G 蛋白家族 G(5)、G(i)、G(q) 和 G(12) 参与与受体特异性信号转导途径激活的联系。最近对 G(12) 蛋白进行了研究，在被 GPCR 激活后，它们介导 RhoGTP 酶鸟嘌呤核苷酸交换因子 (RhoGEF) 的激活，进而激活小 GTP 酶 RhoA。 RhoA 参与许多细胞和生理方面，G(12/13)-Rho 通路功能障碍可导致高血压、心血管疾病、中风、伤口愈合和免疫细胞功能受损、癌症进展和转移或哮喘。在这项研究中，含有增强型绿色荧光蛋白（EGFP）的 RhoGEF 被标记，以检测它们在受体激活后的亚细胞定位和易位。 Constitutively active G alpha(12) and G alpha(13) mutants induced redistribution of these RhoGEFs from the cytosol to the plasma membrane.此外，在以与细胞类型无关的方式激活多个 G(12/13) 偶联 GPCR 后，观察到 p115-RhoGEF 从细胞质到质膜的显着且快速的易位。由 GPCR 激动剂刺激的 p115-RhoGEF 的质膜易位可以通过随后应用相应 GPCR 的拮抗剂来完全且快速地逆转，即 p115-RhoGEF 重新定位回细胞质。 G(12/13) 连接的 GPCR 引起的 RhoGEF 易位可以被量化，因此可用于该途径的药理学研究，并发现 G(12/13) 相关疾病背景中的活性化合物。 J.细胞。生物化学。 104: 1660-1670, 2008。(C) 2008 Wiley-Liss, Inc.
G protein-coupled receptors (GPCRs) are important targets for medicinal agents. Four different G protein families, G(5), G(i), G(q), and G(12), engage in their linkage to activation of receptor-specific signal transduction pathways. G(12) proteins were more recently studied, and upon activation by GPCRs they mediate activation of RhoGTPase guanine nucleotide exchange factors (RhoGEFs), which in turn activate the small GTPase RhoA. RhoA is involved in many cellular and physiological aspects, and a dysfunction of the G(12/13)-Rho pathway can lead to hypertension, cardiovascular diseases, stroke, impaired wound healing and immune cell functions, cancer progression and metastasis, or asthma. In this study, regulator of G protein signaling (RGS) domain-containing RhoGEFs were tagged with enhanced green fluorescent protein (EGFP) to detect their subcellular localization and translocation upon receptor activation. Constitutively active G alpha(12) and G alpha(13) mutants induced redistribution of these RhoGEFs from the cytosol to the plasma membrane. Furthermore, a pronounced and rapid translocation of p115-RhoGEF from the cytosol to the plasma membrane was observed upon activation of several G(12/13)-coupled GPCRs in a cell type-independent fashion. Plasma membrane translocation of p115-RhoGEF Stimulated by a GPCR agonist could be completely and rapidly reversed by subsequent application of an antagonist for the respective GPCR, that is, p115-RhoGEF relocated back to the cytosol. The translocation of RhoGEF by G(12/13)-linked GPCRs can be quantified and therefore used for pharmacological studies of the pathway, and to discover active compounds in a G(12/13)-related disease context. J. Cell. Biochem. 104: 1660-1670, 2008. (C) 2008 Wiley-Liss, Inc.