IDENTIFICATION OF A LIGAND-BINDING SITE OF THE HUMAN ENDOTHELIN-A RECEPTOR AND SPECIFIC REGIONS REQUIRED FOR LIGAND SELECTIVITY

被引:27
作者
ADACHI, M [1 ]
FURUICHI, Y [1 ]
MIYAMOTO, C [1 ]
机构
[1] NIPPON ROCHE RES CTR,DEPT MOLEC GENET,KAMAKURA,KANAGAWA 247,JAPAN
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1994年 / 220卷 / 01期
关键词
D O I
10.1111/j.1432-1033.1994.tb18596.x
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
To investigate the ligand-binding site of the human endothelin-A-receptor subtype (ET(A)), we have produced various chimeric and mutated receptors in chinese hamster ovary cells. The substitution of Lys140 with Ile located in the C-terminus of the second transmembrane region caused a 13-fold reduction in affinity for endothelin-1 (ET-1) and 3.6-fold lower B-max than those values for the original receptor. Correspondingly, the mutated ET(A) receptor with the Lys140-->Ile substitution failed to induce an increase in the intracellular calcium concentration in the presence of 1 nM ET-1. Thus, the Lys140 in the ET(A) receptor is important in ligand binding. ET(A) and ET(B) receptors possess the ET isopeptides selective and non-selective binding activities, respectively Displacement experiments and the binding of I-125-ET-3 to various chimera receptors demonstrated that both the third and fourth extracellular regions, including the flanking transmenbrane regions, are responsible for the ligand-binding selectivity of the ET(A) receptor.
引用
收藏
页码:37 / 43
页数:7
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