人组胺H1受体的基因组克隆、异源表达及药理学特性研究
Genomic cloning, heterologous expression and pharmacological characterization of a human histamine H1 receptor.
作者信息
De Backer M D, Gommeren W, Moereels H, Nobels G, Van Gompel P, Leysen J E, Luyten W H
机构信息
Department of Biochemical Pharmacology, Janssen Research Foundation, Beerse, Belgium.
出版信息
Biochem Biophys Res Commun. 1993 Dec 30;197(3):1601-8. doi: 10.1006/bbrc.1993.2662.
A human histamine H1 receptor gene lacking introns was isolated by screening a human genomic library with a bovine histamine H1 receptor probe. The deduced protein of 487 amino acids showed characteristic properties of G-protein-coupled receptors. The coding region was subcloned into the expression vector pSVL (Pharmacia), and the resulting construct transfected into COS-7 cells. Binding studies with [3H]pyrilamine on membranes from transfected cells revealed saturable specific binding with a KD of 1.2 nM and a Bmax of 3400 fmol/mg protein. Binding affinities of histamine and known histamine antagonists were similar to those for histamine H1 receptors in guinea-pig cerebellum. In transfected COS-7 cells, histamine induced inositol phosphate formation, that was inhibitable by pyrilamine.
通过用牛组胺H1受体探针筛选人基因组文库，分离出一个不含内含子的人组胺H1受体基因。推导的487个氨基酸的蛋白质显示出G蛋白偶联受体的特征特性。将编码区亚克隆到表达载体pSVL（Pharmacia）中，并将所得构建体转染到COS-7细胞中。用[3H]吡苄明对转染细胞的膜进行结合研究，结果显示出可饱和的特异性结合，KD为1.2 nM，Bmax为3400 fmol/mg蛋白质。组胺和已知组胺拮抗剂的结合亲和力与豚鼠小脑组胺H1受体的结合亲和力相似。在转染的COS-7细胞中，组胺诱导肌醇磷酸形成，这可被吡苄明抑制。
Zotero 插件