The Golgi matrix protein GM130:: a specific interacting partner of the small GTPase rab1b
The Golgi matrix protein GM130:: a specific interacting partner of the small GTPase rab1b
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DOI：
10.1093/embo-reports/kve065
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发表时间：
2001-04-01
期刊：
影响因子：
7.7
通讯作者：
Barnekow, A
中科院分区：
文献类型：
--
作者：
Weide, T;Bayer, M;Barnekow, A
为了检测特定的合作伙伴的小高尔基体本地化的GTribrab 1b，我们产生rab 1b突变体，并使用它们作为诱饵蛋白在酵母双杂交筛选。我们分离出了几个特定的相互作用的crons。其中两个编码与大鼠GM 130和人Golgin 95高度同源的大蛋白片段。克隆了全长人GM 130 cDNA，并通过酵母双杂交和体外结合试验详细研究了其与rab 1b的相互作用。在这里，我们报告的rab 1b蛋白的相互作用，特异性与GM 130在一个GTP依赖的方式，因此需要的N-和C-末端的高变区的第一次。我们绘制了GM 130的rab 1b结合位点，并提供了证据表明它不同于先前描述的GM 130蛋白的p115和Grasp 65结合位点。
To detect specific partners of the small Golgi-localized GTPase rab1b we generated rab1b mutants and used them as bait proteins in yeast two-hybrid screens. We isolated several specifically interacting crones. Two of them encode large protein fragments highly homologous to rat GM130 and to human Golgin95. The full-length human GM130 cDNA was cloned and its interaction with rab1b was characterized in detail by yeast two-hybrid and in vitro binding assays. Here we report for the first time that the rab1b protein interacts specificially with GM130 in a GTP-dependent manner and therefore needs the hypervariable regions of the N- and C-termini. We mapped the rab1b binding site of GM130 and provide evidence that it is different to the previously described p115 and Grasp65 binding sites of the GM130 protein.