Hess J F, Borkowski J A, Macneil T, Stonesifer G Y, Fraher J, Strader C D, Ransom R W
Department of Molecular Pharmacology and Biochemistry, Merck Research Laboratories, Rahway, NJ 07065.
Mol Pharmacol. 1994 Jan;45(1):1-8.
The pharmacology of cloned B2 bradykinin receptors heterologously expressed in cell lines lacking any endogenous bradykinin receptors was analyzed. The possibility of B2 bradykinin receptor heterogeneity had been proposed on the basis of numerous studies in various tissue preparations. The results reported here permit a direct evaluation of some of these hypotheses by examining the pharmacological properties of cloned bradykinin receptors. A cloned human B2 bradykinin receptor was stably transfected into Chinese hamster ovary cells. The data suggest that in response to bradykinin (BK), the cloned receptor activates both phosphatidylinositol hydrolysis and arachidonic acid release by independent pathways. Thus, the activation of these two second messenger pathways does not require the existence of two B2 bradykinin receptor subtypes. A mouse gene encoding the B2 bradykinin receptor was isolated, and the coding region was expressed in COS-7 cells. This murine receptor exhibited the pharmacological properties of a "classical" B2 bradykinin receptor. A comparison of the pharmacological profiles of cloned human and murine homologs of the B2 bradykinin receptor indicates that both receptors bind agonists with similar properties. However, the two receptors differ dramatically in their affinity for some peptide antagonists. The mouse receptor has a 60- to 80-fold higher affinity for [D-Arg0Hyp3, Thi5,8,D-Phe7]BK and [D-Arg0,Hyp3,D-Phe7]BK than its human homolog. Thus, the species of a bradykinin receptor can have a significant effect on its pharmacology. The cloning, expression, and pharmacological comparison of human and mouse B2 bradykinin receptor genes indicate that some of the previous reports of B2 receptor subtypes can be explained by species differences in a single B2 bradykinin receptor gene.
对在缺乏任何内源性缓激肽受体的细胞系中异源表达的克隆B2缓激肽受体的药理学进行了分析。基于在各种组织制剂中的大量研究,有人提出了B2缓激肽受体异质性的可能性。本文报道的结果通过检查克隆缓激肽受体的药理学特性,对其中一些假设进行了直接评估。将克隆的人B2缓激肽受体稳定转染到中国仓鼠卵巢细胞中。数据表明,对缓激肽(BK)的反应中,克隆受体通过独立途径激活磷脂酰肌醇水解和花生四烯酸释放。因此,这两条第二信使途径的激活并不需要存在两种B2缓激肽受体亚型。分离出编码B2缓激肽受体的小鼠基因,并将编码区在COS-7细胞中表达。该小鼠受体表现出“经典”B2缓激肽受体的药理学特性。对B2缓激肽受体的克隆人同源物和小鼠同源物的药理学特征进行比较表明,两种受体结合具有相似特性的激动剂。然而,这两种受体对某些肽拮抗剂的亲和力差异很大。小鼠受体对[D-Arg0Hyp3,Thi5,8,D-Phe7]BK和[D-Arg0,Hyp3,D-Phe7]BK的亲和力比其人类同源物高60至80倍。因此,缓激肽受体的物种对其药理学有显著影响。人及小鼠B2缓激肽受体基因的克隆、表达及药理学比较表明,先前关于B2受体亚型的一些报道可以用单一B2缓激肽受体基因的物种差异来解释。