Characterization of a human cDNA that encodes a functional receptor for platelet activating factor.
Characterization of a human cDNA that encodes a functional receptor for platelet activating factor.
复制标题
编码血小板激活因子功能受体的人类 cDNA 的表征。
DOI：
10.1016/s0006-291x(05)81261-6
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发表时间：
1991
影响因子：
3.1
通讯作者：
Cochrane,CG
中科院分区：
文献类型：
--
作者：
Ye,RD;Prossnitz,ER;Zou,AH;Cochrane,CG
我们用基于视紫红质型受体保守序列的简并寡核苷酸探针筛选HL-60粒细胞cDNA文库，克隆了血小板活化因子（PAF）受体的cDNA。该342个氨基酸的受体含有7个推定的跨膜结构域，但在N-末端缺乏用于N-连接糖基化的位点。表达克隆cDNA的稳定转染成纤维细胞对亚纳摩尔PAF刺激产生钙动员反应，这可以被PAF拮抗剂L 659，989抑制。在人胎盘、肺和分化的HL-60粒细胞中检测到PAF受体信息为约4kb的单一种类。克隆的cDNA在缺乏内源性PAF受体的未分化HL-60细胞中的表达导致PAF拮抗剂WEB 2086的特异性和饱和结合，解离常数为30.7 nM。
We have cloned a cDNA for the platelet activating factor (PAF) receptor by screening an HL-60 granulocyte cDNA library with degenerate oligonucleotide probes based on conserved sequences of rhodopsin-type receptors. The 342-amino acid receptor contains 7 putative transmembrane domains, but lacks sites for N-linked glycosylation at the N-terminus. Stably transfected fibroblasts expressing the cloned cDNA responded to sub-nanomolar PAF stimulation with calcium mobilization, which could be inhibited by the PAF antagonist L659,989. The PAF receptor message was detected as a single species of approximately 4 kb in human placenta, lung, and differentiated HL-60 granulocytes. Expression of the cloned cDNA in undifferentiated HL-60 cells devoid of endogenous PAF receptor resulted in specific and saturable binding of the PAF antagonist WEB 2086 with a dissociation constant of 30.7 nM.