ESTABLISHMENT OF CHINESE-HAMSTER OVARY CELL-LINES STABLY EXPRESSING THE CLONED HUMAN TYPE-1 ANGIOTENSIN-II RECEPTOR AND CHARACTERIZATION OF THE EXPRESSED RECEPTOR

被引:7
作者
ISHIDO, M
KONDOH, M
OHNISHI, J
KOBAYASHI, M
MITSUI, Y
FURUTA, H
GUO, DF
INAGAMI, T
BIRNBAUMER, M
MURAKAMI, K
MIYAZAKI, H
机构
[1] AGCY IND SCI & TECHNOL,FERMENTAT RES INST,DIV CELL SCI & TECHNOL,IBARAKI 305,JAPAN
[2] BAYLOR COLL MED,DEPT CELL BIOL,HOUSTON,TX 77030
[3] VANDERBILT UNIV,MED CTR,SCH MED,DEPT BIOCHEM,NASHVILLE,TN 37232
来源
BIOMEDICAL RESEARCH-TOKYO | 1992年 / 13卷 / 05期
关键词
D O I
10.2220/biomedres.13.349
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
We have isolated the human angiotensin II (AII) type 1 receptor (AT1) gene, whose coding region was contained within a single exon and have established Chinese hamster ovary (CHO) cell lines stably expressing this recombinant receptor. The expressed receptor revealed the typical binding characteristics of the AT1 receptor to several All receptor agonists and antagonists. Treatment of the intact CHO cells with the sulfhydryl reducing reagent dithiothreitol predominantly decreased the radioligand binding, indicating that a disulfhydryl bond(s) necessary for AII binding is located on extracellular domains of the receptor molecule. Addition of AII to the cells evoked a rapid, transient increase in intracellular Ca2+ concentrations followed by a lower, sustained phase; the initial increase was mediated by inositol triphosphate and the second phase was due to Ca2+ influx through voltage-dependent L-type Ca2+ channels. These results demonstrate that the cloned gene indeed encodes a functional AT1 receptor. Furthermore, the appearance of a single hybridization band on genomic Southern blot analysis performed both at high and low stringency, suggests the existence of one copy of the AT1 receptor in the human genome in contrast to that of the two distinct AT1 receptor genes in the rat.
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页码:349 / 356
页数:8
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