首页> 外文期刊>Cellular Signalling >Mutations in the carboxy-terminus of the third intracellular loop of the human recombinant VPAC(1) receptor impair VIP-stimulated [Ca2+](i) increase but not adenylate cyclase stimulation
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Mutations in the carboxy-terminus of the third intracellular loop of the human recombinant VPAC(1) receptor impair VIP-stimulated [Ca2+](i) increase but not adenylate cyclase stimulation

机译:人类重组VPAC(1)受体的第三个细胞内环的羧基末端的突变会损害VIP刺激的[Ca2 +](i)增加,但不会腺苷酸环化酶刺激

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摘要

The vasoactive intestinal polypeptide (VIP) VPAC(1) receptor is preferentially coupled to Galphas protein that stimulates adenylate cyclase activity and also to Galphaq and Galphai proteins that stimulate the inositol phosphate/calcium pathway. Previous studies indicated the importance of the third intracellular loop of the receptor for G protein coupling. By site-directed mutation of the human recombinant receptor expressed in Chinese hamster ovary cells, we identified two domains in this loop that contain clusters of basic residues conserved in most of the G-protein-coupled seven transmembrane domains receptors. We found that mutations in the proximal domain (K-322) reduced the capability of VIP to increase adenylate cyclase activity without any change in the calcium response, whereas mutations in the distal part of the loop (R-338, L-339, R-341) markedly reduced the calcium increase and Galphai coupling but only weakly the adenylate cyclase activity. Thus, the interaction of different G proteins with the VPAC(1) receptor involves different receptor sub-domains. (C) 2004 Elsevier Inc. All rights reserved.
机译:血管活性肠多肽(VIP)VPAC(1)受体优先与刺激腺苷酸环化酶活性的Galphas蛋白偶联,并与刺激肌醇磷酸/钙途径的Galphaq和Galphai蛋白偶联。先前的研究表明该受体的第三细胞内环对于G蛋白偶联的重要性。通过在中国仓鼠卵巢细胞中表达的人类重组受体的定点突变,我们在该环中鉴定了两个结构域,其中包含在大多数G蛋白偶联的七个跨膜结构域受体中保守的基本残基簇。我们发现近端结构域(K-322)中的突变降低了VIP增加腺苷酸环化酶活性的能力,而钙反应无任何变化,而环远端(R-338,L-339,R -341)明显降低了钙的增加和Galphai耦合,但腺苷酸环化酶的活性却很弱。因此,不同的G蛋白与VPAC(1)受体的相互作用涉及不同的受体亚结构域。 (C)2004 Elsevier Inc.保留所有权利。

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