CONTROL OF YEAST MATING SIGNAL TRANSDUCTION BY A MAMMALIAN BETA-2-ADRENERGIC RECEPTOR AND GS ALPHA-SUBUNIT
CONTROL OF YEAST MATING SIGNAL TRANSDUCTION BY A MAMMALIAN BETA-2-ADRENERGIC RECEPTOR AND GS ALPHA-SUBUNIT
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DOI：
10.1126/science.2171146
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发表时间：
1990-10-05
期刊：
影响因子：
56.9
通讯作者：
LEFKOWITZ, RJ
中科院分区：
文献类型：
--
作者：
KING, K;DOHLMAN, HG;LEFKOWITZ, RJ
为了促进受体-G蛋白相互作用的功能和机制研究，通过在酿酒酵母中表达人β2-肾上腺素能受体(h.β-AR)。这是通过将修饰的 h.beta.-AR 基因置于半乳糖诱导型 GAL1 启动子的控制下来实现的。经过半乳糖诱导后，功能性 h.β-AR 的表达浓度是任何人体组织中的浓度的数百倍。根据竞争性配体结合实验确定，酵母中表达的h.β-AR表现出特有的亲和力、特异性和立体选择性。在共表达h.β-AR和哺乳动物G蛋白(Gs)α的细胞中实现了β-肾上腺素能受体激动剂对酵母信息素应答途径的部分激活。 subunit.sbd.证明这些组件可以相互耦合，并在以年份表示时与下游效应器耦合。这种体内重建系统提供了一种检查配体结合和 G 蛋白与细胞表面受体偶联的新方法。
To facilitate functional and mechanistic studies of receptor-G protein interactions by expression of the human .beta.2-adrenergic receptor (h.beta.-AR) has been expressed in Saccharomyces cerevisiae. This was achieved by placing a modified h.beta.-AR gene under control of the galactose-inducible GAL1 promoter. After induction by galactose, functional h.beta.-AR was expressed at a concentration several hundred times as great as that found in any human tissue. As determined from competitive ligand binding experiments, h.beta.-AR expressed in yeast displayed characteristic affinities, specificity, and stereoselectivity. Partial activation of the yeast pheromone response pathway by .beta.-adrenergic receptor agonists was achieved in cells coexpressing h.beta.-AR and a mammalian G protein (Gs) .alpha. subunit.sbd.demonstrating that these components can couple to each other and to downstream effectors when expressed in year. This in vivo reconstitution system provides a new approach for examining ligand binding and G protein coupling to cell surface receptors.