Lai J, Mei L, Roeske W R, Chung F Z, Yamamura H I, Venter J C
Section of Receptor Biochemistry and Molecular Biology, National Institute of Neurologic, Communicative Disorders and Stroke, Bethesda, MD 20892.
Life Sci. 1988;42(24):2489-502. doi: 10.1016/0024-3205(88)90348-7.
A rat genomic DNA clone was isolated by its homology with a conserved primary sequence among the mammalian and avian beta adrenergic and porcine muscarinic receptors. A gene identified in this clone was highly homologous to the rat M1 muscarinic receptor. Stable expression of this gene was achieved in an established murine fibroblast cell line, B82. The gene product exhibits M1 type muscarinic receptor characteristics, as it has high affinity for PZ but low affinity for AF-DX 116. Carbachol stimulated the hydrolysis of phosphatidylinositols in the transfected cells. Pirenzepine had a more potent inhibitory effect on this response than AF-DX 116 since their functional inhibition constants were 13 nM and 480 nM, respectively, which is consistent with an M1 pharmacological profile. These data suggest that the M1 muscarinic receptor encoded by the gene is coupled to the hydrolysis of phosphatidylinositols after transfecting this gene into the B82 cells.
通过与哺乳动物和禽类β肾上腺素能受体及猪毒蕈碱受体中保守的一级序列具有同源性,分离出一个大鼠基因组DNA克隆。在该克隆中鉴定出的一个基因与大鼠M1毒蕈碱受体高度同源。在已建立的小鼠成纤维细胞系B82中实现了该基因的稳定表达。该基因产物表现出M1型毒蕈碱受体特征,因为它对PZ具有高亲和力而对AF-DX 116具有低亲和力。卡巴胆碱刺激转染细胞中磷脂酰肌醇的水解。哌仑西平对该反应的抑制作用比AF-DX 116更强,因为它们的功能抑制常数分别为13 nM和480 nM,这与M1药理学特征一致。这些数据表明,将该基因转染到B82细胞后,由该基因编码的M1毒蕈碱受体与磷脂酰肌醇的水解偶联。