Molecular cloning and functional expression of the guinea pig α1a-adrenoceptor
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- 作者：Gonzá ; lez-Espinosa ; Claudia ; Romero-Á ; vila ; Marí ; a Teresa ; Mora-Rodrí ; guez ; Dulce Marí ; a ; et. al.
- 关键词：α ; 1-Adrenoceptor ; α ; 1a-Adrenoceptor ; Cloning
- 刊名：European Journal of Pharmacology
- 出版年：2001
- 出版时间：August 31, 2001
- 年：2001
- 卷：426
- 期：3
- 页码：147-155
- 全文大小：331 K
文摘
In the present paper, the cloning and expression of the guinea pig α1A-adrenoceptor is presented. The nucleotide sequence had an open reading frame of 1401 bp that encoded a 466 amino-acid protein with an estimated molecular mass of 
51.5 kDa. When the clone was expressed in Cos-1 cells, specific high-affinity binding of [3H]prazosin and [3H]tamsulosin was observed. Chloroethylclonidine treatment of membranes slightly decreased the total binding with both radioligands. Binding competition experiments using [3H]tamsulosin showed the following potency order: (a) for agonists: oxymetazoline
epinephrine>norepinephrine>methoxamine, and (b) for antagonists: prazosin≥5-methyl-urapidil=benoxathian>phentolamineBMY 7378 (8-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-8-azaspiro[4,5]decane-7,9-dione). Photoaffinity labeling using [125I-aryl]azido-prazosin revealed a major broad band with a molecular mass between 70 and 80 kDa. The receptor was functional, as evidenced by an epinephrine-increased production of [3H]inositol phosphates that was blocked by prazosin.
51.5 kDa. When the clone was expressed in Cos-1 cells, specific high-affinity binding of [3H]prazosin and [3H]tamsulosin was observed. Chloroethylclonidine treatment of membranes slightly decreased the total binding with both radioligands. Binding competition experiments using [3H]tamsulosin showed the following potency order: (a) for agonists: oxymetazolineepinephrine>norepinephrine>methoxamine, and (b) for antagonists: prazosin≥5-methyl-urapidil=benoxathian>phentolamineBMY 7378 (8-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-8-azaspiro[4,5]decane-7,9-dione). Photoaffinity labeling using [125I-aryl]azido-prazosin revealed a major broad band with a molecular mass between 70 and 80 kDa. The receptor was functional, as evidenced by an epinephrine-increased production of [3H]inositol phosphates that was blocked by prazosin.