应用酵母双杂交研究G蛋白通路中的蛋白因子

目的 探寻G蛋白信号传导途径中与Gβ亚基相互作用下的游蛋白因子,揭示G蛋白信号传导通路的机制。方法 采用敏感性较高的酵母双杂交体系,构建含Gβ亚基基因的饵质粒筛选人脑cDNA文库。结果 获得了肌动蛋白集束调控蛋白（actin bundling protein）的编码基因和两个新基因片段,GeneBank登录号分别为288405(427bp)及AF288406＊2832BP）。结论 提示在人脑组织中Gβ亚基作为结构和功能单位可能通过与actin bundling protein及另两种未知的蛋白因子间的相互作用而介导了信号的传导,对于揭示G蛋白信号传递通路与actin细胞骨架之间的关系起重要的提示作用。

Hunting for novel protein factors in G-protein pathway with yeast two-hybrid system

OBJECTIVE: To explore the protein factors that could interact with G beta subunit within the G protein signal transducing pathway. METHODS: The highly sensitive protein-protein interaction system--Yeast Two-hybrid System was applied to screen the human cDNA library with constructed "Bite plasmid" containing G beta subunit gene fragment. And then the false positive test was adapted. RESULTS: Three positive gene fragments were obtained. One codes for "Actin bundling protein". The other two are new ones and their Gene Bank accession numbers are AF288405 and AF288406 respectively. CONCLUSIONS: It is the first time to find that among human brain tissue, G beta subunit becomes a structural or functional unit interacts with actin bundling protein and the other two unknown protein factors to activate the following pathway. This result may be important to understand the relationship between the actin cytoskeleton and G proteins.