Recombinant antibodies to the small GTPase Rab6 as conformation sensors
Recombinant antibodies to the small GTPase Rab6 as conformation sensors
复制标题
DOI：
10.1126/science.1083911
复制
发表时间：
2003-05-09
期刊：
影响因子：
56.9
通讯作者：
Perez, F
中科院分区：
文献类型：
--
作者：
Nizak, C;Monier, S;Perez, F
在这里，我们报告的方法，抗体噬菌体展示的基础上，产生分子构象传感器。产生特异于小的鸟苷三磷酸酶（GTIPs）Rab6（膜交通的调节剂）的鸟苷三磷酸（GTP）结合构象的重组抗体，并用于在固定的细胞中定位Rab6.GTP，并且在绿色荧光蛋白（GFP）标记和细胞内表达之后，在体内跟踪Rab6.GTP。Rab6在高尔基体和转运中间体上处于GTP结合构象，转运中间体的几何形状受Rab6活性的调节。更一般地，相同的方法可以应用于可以在体外锁定在特定构象的其他分子。
Here we report an approach, based on antibody phage display, to generate molecular conformation sensors. Recombinant antibodies specific to the guanosine triphosphate (GTP)-bound conformation of the small guanosine triphosphatase (GTPase) Rab6, a regulator of membrane traffic, were generated and used to locate Rab6.GTP in fixed cells, and, after green fluorescent protein (GFP) tagging and intracellular expression, to follow Rab6.GTP in vivo. Rab6 was in its GTP-bound conformation on the Golgi apparatus and transport intermediates, and the geometry of transport intermediates was modulated by Rab6 activity. More generally, the same approach could be applied to other molecules that can be locked in a particular conformation in vitro.