Guillén A, Jallon J M, Fehrentz J A, Pantaloni C, Bockaert J, Homburger V
Centre CNRS-INSERM de Pharmacologie-Endocrinologie, Montpellier, France.
EMBO J. 1990 May;9(5):1449-55. doi: 10.1002/j.1460-2075.1990.tb08261.x.
G proteins couple receptors for extracellular signals to several intracellular effector systems and play a key role in signalling transduction mechanisms. In particulate preparations of Drosophila melanogaster heads, only one substrate for pertussis toxin at 39-40 kd was detected. This substrate, which showed only one isoform when analysed by isoelectric focusing, was recognized by immunoblotting and immunoprecipitation techniques using a polyclonal antibody against the alpha subunit of the Go protein purified from bovine brain and can be thus considered as a Go-like protein. Antibodies obtained against a carboxy-terminal sequence of the alpha subunit of Go (but not of Gi1 or Gi2) and against an internal sequence shared by all the alpha subunits, were also able to cross-react with the alpha subunit of this protein in insects. We have also studied the Go-like protein in several D.melanogaster mutants, primarily in memory and learning mutants. In these mutants there was a sex-dependent enhancement in pertussis toxin-catalysed ADP-ribosylation with respect to the wild-type. This increase could be attributed in part to an increase in the alpha subunit of the Go-like protein, as revealed by immunoblotting with anti-Go alpha polyclonal antibody. This report constitutes the first evidence for the participation of a Go protein in learning and memory.
G蛋白将细胞外信号受体与多种细胞内效应系统偶联起来,在信号转导机制中起关键作用。在黑腹果蝇头部的微粒体制备物中,仅检测到一种39 - 40kd的百日咳毒素底物。通过等电聚焦分析时,该底物仅显示一种同工型,使用针对从牛脑纯化的Go蛋白α亚基的多克隆抗体,通过免疫印迹和免疫沉淀技术可识别该底物,因此可将其视为一种类Go蛋白。针对Go蛋白α亚基的羧基末端序列(而非Gi1或Gi2的羧基末端序列)以及所有α亚基共有的内部序列产生的抗体,也能够与昆虫体内这种蛋白质的α亚基发生交叉反应。我们还研究了几种黑腹果蝇突变体中的类Go蛋白,主要是记忆和学习突变体。在这些突变体中,相对于野生型,百日咳毒素催化的ADP - 核糖基化存在性别依赖性增强。如用抗Goα多克隆抗体进行免疫印迹所显示,这种增加部分可归因于类Go蛋白α亚基的增加。本报告构成了Go蛋白参与学习和记忆的首个证据。