Ullrich O, Stenmark H, Alexandrov K, Huber L A, Kaibuchi K, Sasaki T, Takai Y, Zerial M
European Molecular Biology Laboratory, Heidelberg, Federal Republic of Germany.
J Biol Chem. 1993 Aug 25;268(24):18143-50.
Rab proteins comprise a family of small GTPases that serve a regulatory role in membrane traffic. These proteins are in part cytosolic and in part associated with the membranes of specific exocytic and endocytic organelles. Smg p25A/rab3A GDI, a cytosolic protein which inhibits the dissociation of GDP from smg p25A/rab3A, Sec4p, and rab11, has also been found to prevent association of rab3A with the membrane. In this study, we have used Madin-Darby canine kidney cells permeabilized with the bacterial toxin streptolysin O to test the general activity of rab3A GDI in modulating the membrane association of various small GTP-binding proteins. Rab3A GDP dissociation inhibitor (GDI) removed from the membrane all rab proteins we have tested and inhibited the membrane binding of in vitro translated rab proteins. However, rab3A GDI had a limited effect on the membrane association of a mutant rab5 protein which contained a farnesylated cysteine motif. Finally, we found that, although rab3A GDI resides primarily in the cytosol, it is also associated with compartments of the exocytic and endocytic pathways. Since rab3A GDI can modulate the membrane association of various rab proteins, we propose to rename it rab GDI.
Rab蛋白构成了一个小GTP酶家族,在膜运输中起调节作用。这些蛋白部分存在于细胞质中,部分与特定的胞吐和胞吞细胞器的膜相关联。Smg p25A/rab3A GDI是一种细胞质蛋白,可抑制GDP从smg p25A/rab3A、Sec4p和rab11上解离,也已被发现可阻止rab3A与膜的结合。在本研究中,我们使用经细菌毒素链球菌溶血素O通透处理的Madin-Darby犬肾细胞,来测试rab3A GDI在调节各种小GTP结合蛋白的膜结合方面的一般活性。Rab3A GDP解离抑制剂(GDI)从膜上移除了我们测试的所有rab蛋白,并抑制了体外翻译的rab蛋白与膜的结合。然而,rab3A GDI对含有法尼基化半胱氨酸基序的突变型rab5蛋白的膜结合影响有限。最后,我们发现,尽管rab3A GDI主要存在于细胞质中,但它也与胞吐和胞吞途径的区室相关联。由于rab3A GDI可以调节各种rab蛋白的膜结合,我们建议将其重新命名为rab GDI。