Shapiro R A, Wakimoto B T, Subers E M, Nathanson N M
Department of Pharmacology, University of Washington, Seattle 98195.
Proc Natl Acad Sci U S A. 1989 Nov;86(22):9039-43. doi: 10.1073/pnas.86.22.9039.
Genomic and cDNA clones encoding a muscarinic acetylcholine receptor from Drosophila melanogaster have been isolated. Sequence analysis demonstrates that this gene encodes a receptor with a high degree of amino acid identity to the mammalian muscarinic acetylcholine receptors and has three introns in the portion of the gene encoding the third putative cytoplasmic loop. A full-length cDNA clone has been placed under the control of the mouse metallothionein promotor and transfected into mouse Y1 adrenal cells. The receptor expressed in these cells exhibits the high-affinity binding for the antagonists quinuclidinyl benzilate and atropine expected of a muscarinic receptor. The Drosophila muscarinic receptor, when expressed in Y1 cells, is physiologically active, as measured by agonist-dependent stimulation of phosphatidylinositol metabolism.
编码来自黑腹果蝇毒蕈碱型乙酰胆碱受体的基因组和cDNA克隆已被分离出来。序列分析表明,该基因编码的受体与哺乳动物毒蕈碱型乙酰胆碱受体具有高度的氨基酸同一性,并且在编码第三个推定细胞质环的基因部分中有三个内含子。一个全长cDNA克隆已置于小鼠金属硫蛋白启动子的控制下,并转染到小鼠Y1肾上腺细胞中。在这些细胞中表达的受体对毒蕈碱受体拮抗剂喹宁环基苯甲酸酯和阿托品表现出预期的高亲和力结合。当在Y1细胞中表达时,果蝇毒蕈碱受体具有生理活性,这通过激动剂依赖性的磷脂酰肌醇代谢刺激来衡量。