FUNCTIONAL EXPRESSION OF RECOMBINANT HUMAN LUTEINIZING HUMAN CHORIOGONADOTROPIN RECEPTOR

被引:9
作者
IGARASHI, S
MINEGISHI, T
NAKAMURA, K
NAKAMURA, M
TANO, M
MIYAMOTO, K
IBUKI, Y
机构
[1] GUNMA UNIV,SCH MED,DEPT OBSTET & GYNECOL,MAEBASHI,GUMMA 371,JAPAN
[2] NATL CARDIOVASC CTR,RES INST,SUITA,OSAKA 565,JAPAN
关键词
D O I
10.1006/bbrc.1994.1696
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The functional capacity of the recombinant human LH/hCG receptor was tested on the basis of gonadotropin stimulation of cAMP production by stable transfections of Chinese hamster ovary (CHO) cells. A CHO cell line expressed with the hLH/hCG receptor cDNA covering the entire amino acid coding region revealed the presence of LH/hCG binding site (Kd:1.45x10(-10)M) on the plasma membrane. Treatment of transfected cells(CHO-LH/hCGR) with hCG induced dose-dependent increases in intracellular cAMP production, indicating that the expressed human LH/hCG receptor functionally couples with endogenous adenylyl cyclase. Although hCG induced dose-dependent increases in cAMP production. rat and bovine LH and human FSH did not alter cAMP levels compared to control values. Northern blot analysis with a cRNA probe derived from human LH/hCG receptor cDNA indicated the presence of three LH/hCG receptor mRNA transcripts(5.4, 3.6 and 2.4 kilobases) in RNA prepared from human ovary. Preincubation of CHO-LH/hCGR cells with hCG for 16h decreased the subsequent cAMP production caused by a 30min results indicate that desensitization to hCG stimulation occurs in CHO-LH/hCGR cells. Therefore, this cell line provides a tool with which to pursue detailed studies on the molecular basis of LH/hCG induced desensitization. (C) 1994 Academic Press, Inc.
引用
收藏
页码:248 / 256
页数:9
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