EXPRESSION CLONING OF A RAT B2-BRADYKININ RECEPTOR
EXPRESSION CLONING OF A RAT B2-BRADYKININ RECEPTOR
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DOI：
10.1073/pnas.88.17.7724
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发表时间：
1991-09-01
影响因子：
11.1
通讯作者：
JARNAGIN, K
中科院分区：
文献类型：
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作者：
MCEACHERN, AE;SHELTON, ER;JARNAGIN, K
采用克隆选择策略，从大鼠子宫文库中分离到一个编码功能性缓激肽受体的基因，并利用非洲爪哇的卵母细胞来检测缓激肽反应的表达。预测的蛋白质与七个跨膜G蛋白偶联受体超家族同源。缓激肽及其类似物在表达受体的卵母细胞上以效力顺序刺激氯电流：缓激肽几乎等于Lys-缓激肽>；[Tyr8]缓激肽。这是在与大鼠子宫可溶性受体的竞争结合试验中观察到的这些化合物的效力等级顺序。DES-Arg9-Bradykinin(10-MU-M)对表达该受体的卵母细胞无反应；(HUS，该基因编码B2型缓激肽受体。[Thi5，8，DPhe7][Thi5，8，DPhe7]缓激肽是一种非常弱的部分激动剂，可以抑制缓激肽介导的离子通量，提示该基因编码的是一种平滑肌而不是神经元的B2受体亚型。受体信息的分布与先前报道的缓激肽功能和/或结合在几个组织中一致，并在大鼠子宫、输精管、肾、肺、心脏、回肠、睾丸和脑中发现。受体亚型是一种可能性，因为几个组织包含两个或三个信息种类(4.0、5.7和6.5千碱基)。Southern印迹高严格率分析表明，大鼠、豚鼠和人类基因组中只有一个基因。由于缓激肽是疼痛的Kev介体，了解该受体的一级结构将有助于对该受体的分子理解，并有助于设计用于止痛的拮抗剂。
A cDNA encoding a functional bradykinin receptor was isolated from a rat uterus library by a clonal selection strategy using Xenopus laevis oocytes to assay for expression of bradykinin responses. The predicted protein is homologous to the seven transmembrane G protein-coupled superfamily of receptors. Bradykinin and its analogs stimulate a Cl- current in oocytes expressing the receptor with the rank order of potency: bradykinin almost-equal-to Lys-bradykinin > [Tyr8]bradykinin >> [Phe6]bradykinin. This is the rank order of potency observed for these compounds in competitive binding assays on soluble receptor from rat uterus. Des-Arg9-bradykinin (10-mu-M) elicits no response when applied to oocytes expressing the receptor; (hus, the cDNA encodes a B2 type bradykinin receptor. [Thi5,8,DPhe7]bradykinin, where Thi is beta-(2-thienyl)-alanine, is a very weak partial agonist and inhibits the bradykinin-mediated ion flux, suggesting the cDNA encodes a smooth muscle, rather than a neuronal, B2 receptor subtype. Receptor message has a distribution consistent with previous reports of bradykinin function and/or binding in several tissues and is found in rat uterus, vas deferens, kidney, lung, heart, ileum, testis, and brain. Receptor subtypes are a possibility because several tissues contain two or three message species (4.0, 5.7, and 6.5 kilobases). Southern blot high-stringency analysis demonstrated that the rat, guinea pig, and human genomes contain a single gene. As bradykinin is a kev mediator of pain, knowledge of the primary structure of this receptor will allow a molecular understanding of the receptor and aid the design of antagonists for pain relief.