从大鼠垂体和小鼠垂体肿瘤细胞系克隆的促性腺激素释放激素（GnRH）受体的分子与功能特性

Molecular and functional characterization of GnRH receptors cloned from rat pituitary and a mouse pituitary tumor cell line.

作者信息

Perrin M H, Bilezikjian L M, Hoeger C, Donaldson C J, Rivier J, Haas Y, Vale W W

机构信息

Clayton Foundation Laboratories for Peptide Biology, Salk Institute for Biological Studies, La Jolla, CA 92037.

出版信息

Biochem Biophys Res Commun. 1993 Mar 31;191(3):1139-44. doi: 10.1006/bbrc.1993.1335.

DOI:10.1006/bbrc.1993.1335

PMID:7916600

Abstract

A cDNA for a GnRH receptor (mtGnRH-R) was obtained from a mouse gonadotropic pituitary cell line (alpha T3-1) by expression cloning. This full-length cDNA was subsequently used as a probe to clone a rat pituitary GnRH receptor (rGnRH-R). The two receptors differ by 13 amino acids and are 100% identical to those recently reported. The analysis of the cloned receptors by photoaffinity-labeling followed by SDS-PAGE reveals a major band of approximately 70 kDa. This is in contrast to the native rat pituitary and mouse alpha T3-1 receptors whose major labeled species migrate with an apparent size of approximately 45 kDa. Functional studies reveal that both receptors, when transiently expressed in COSM6 cells, can bind GnRH with high affinity and transduce the stimulation of IP3 accumulation in response to GnRH.

摘要

通过表达克隆从小鼠促性腺垂体细胞系（αT3 - 1）中获得了促性腺激素释放激素受体（mtGnRH - R）的互补DNA（cDNA）。该全长cDNA随后被用作探针来克隆大鼠垂体促性腺激素释放激素受体（rGnRH - R）。这两种受体有13个氨基酸不同，并且与最近报道的受体100%相同。通过光亲和标记后进行十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE）对克隆的受体进行分析，显示出一条约70 kDa的主要条带。这与天然大鼠垂体和小鼠αT3 - 1受体形成对比，其主要标记物种的迁移表观大小约为45 kDa。功能研究表明，当在COSM6细胞中瞬时表达时，这两种受体都能以高亲和力结合促性腺激素释放激素（GnRH），并转导GnRH刺激引起的肌醇三磷酸（IP3）积累。