Molecular and functional characterization of GnRH receptors cloned from rat pituitary and a mouse pituitary tumor cell line.
Molecular and functional characterization of GnRH receptors cloned from rat pituitary and a mouse pituitary tumor cell line.
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DOI：
10.1006/bbrc.1993.1335
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发表时间：
1993-03
影响因子：
3.1
通讯作者：
M. Perrin;L. Bilezikjian;C. Hoeger;Cynthia J. Donaldson;J. Rivier;Yaira Haas;W. Vale
中科院分区：
文献类型：
--
作者：
M. Perrin;L. Bilezikjian;C. Hoeger;Cynthia J. Donaldson;J. Rivier;Yaira Haas;W. Vale
通过表达克隆从小鼠促性腺激素垂体细胞系（α T3-1）获得GnRH受体（mtGnRH-R）的cDNA。该全长cDNA随后被用作探针以克隆大鼠垂体GnRH受体（rGnRH-R）。这两种受体相差13个氨基酸，与最近报道的受体100%相同。通过光亲和标记和SDS-PAGE对克隆受体进行分析，显示出约70 kDa的主带。这与天然大鼠垂体和小鼠α T3-1受体相反，其主要标记物质以约45 kDa的表观大小迁移。功能研究表明，这两种受体在COSM 6细胞中瞬时表达时，可以高亲和力结合GnRH，并抑制GnRH对IP 3积累的刺激。
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.