Lang J, Costa T
Department of Neuropharmacology, Max-Planck-Institut für Psychiatrie, Planegg-Martinsried, F.R.G.
Biochem Biophys Res Commun. 1987 Oct 29;148(2):838-48. doi: 10.1016/0006-291x(87)90952-1.
To obtain antisera specific for the GTP-binding protein Gi alpha we immunized rabbits against a synthetic peptide derived from the N-terminal (3-17) sequence predicted from the rat Gi alpha cDNA clone published by Itoh et al. (1986) (Proc. Natl. Acad. Sci. USA 83, 3776-3780). Western-blot analysis of bovine brain G-proteins purified and resolved by hydrophobic chromatography and of rat striatal membranes, indicate that this antiserum does not recognize 41 kDa alpha i or 39 kDa alpha o. However, it reacts with a 40 kDa alpha-subunit. The data suggest that the sequence deduced from the rat G alpha i cDNA corresponds to a G40 alpha protein and that N-terminus directed antisera are useful tools to discriminate between two different G alpha i-like types of G-proteins present in mammalian brain.
为了获得针对GTP结合蛋白Giα的特异性抗血清,我们用一种合成肽免疫兔子,该合成肽来源于Itoh等人(1986年)发表的大鼠Giα cDNA克隆预测的N端(3 - 17)序列(《美国国家科学院院刊》83卷,3776 - 3780页)。对通过疏水色谱法纯化和分离的牛脑G蛋白以及大鼠纹状体膜进行的蛋白质免疫印迹分析表明,这种抗血清不能识别41 kDa的αi或39 kDa的αo。然而,它能与一个40 kDa的α亚基发生反应。数据表明,从大鼠Gαi cDNA推导的序列对应于一种G40α蛋白,并且N端导向的抗血清是区分哺乳动物脑中存在的两种不同类型的Gαi样G蛋白的有用工具。