Jansson C C, Karp M, Oker-Blom C, Näsman J, Savola J M, Akerman K E
Department of Biochemistry and Pharmacy, Abo Akademi University, Turku, Finland.
Eur J Pharmacol. 1995 Jul 18;290(2):75-83. doi: 10.1016/0922-4106(95)90019-5.
The baculovirus expression vector system utilizing the strong polyhedrin gene promoter of the Autographa californica nuclear polyhedrosis virus (AcNPV) was used for high level expression of the two alpha 2-adrenoceptor subtypes alpha 2A-C10 and alpha 2B-C2 in Spodoptera frugiperda (Sf-9) insect cells. For rapid screening of recombinant viruses the luciferase gene was expressed under the early ETL-promoter (early transcript large) in the same plasmid. Both receptor subtypes showed the same rank order of binding affinity for four agonists tested: dexmedetomidine > l-medetomidine = clonidine > noradrenaline. For the alpha 2A-C10 subtype, these agonists inhibited forskolin stimulated cAMP production through pertussis toxin sensitive G-proteins. In contrast, for the alpha 2B-C2 subtype the agonists stimulated both basal and forskolin stimulated cAMP production.
利用苜蓿银纹夜蛾核型多角体病毒(AcNPV)的强多角体蛋白基因启动子的杆状病毒表达载体系统,用于在草地贪夜蛾(Sf-9)昆虫细胞中高水平表达两种α2-肾上腺素能受体亚型α2A-C10和α2B-C2。为了快速筛选重组病毒,荧光素酶基因在同一质粒的早期ETL启动子(早期转录本大)下表达。两种受体亚型对所测试的四种激动剂显示出相同的结合亲和力顺序:右美托咪定>左旋美托咪定=可乐定>去甲肾上腺素。对于α2A-C10亚型,这些激动剂通过百日咳毒素敏感的G蛋白抑制福斯高林刺激的cAMP产生。相比之下,对于α2B-C2亚型,激动剂刺激基础和福斯高林刺激的cAMP产生。