Kinetic activation of Rab8 guanine nucleotide exchange factor Rabin8 by Rab11.
Kinetic activation of Rab8 guanine nucleotide exchange factor Rabin8 by Rab11.
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Rab11 对 Rab8 鸟嘌呤核苷酸交换因子 Rabin8 的动力学激活。
DOI：
10.1007/978-1-4939-2569-8_8
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Guo,Wei
中科院分区：
文献类型：
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作者：
Feng,Shanshan;Wu,Bin;Peränen,Johan;Guo,Wei
小gtpase的Rab家族作为分子开关，控制囊泡运输的各个阶段。Rab8在反式高尔基网络（TGN）的胞外运输和内体到质膜的再循环中起作用。Rabin8是Rab8的主要鸟嘌呤核苷酸交换因子（GEF）。它通过催化Rab8的GDP释放来激活随后的GTP加载。然而，Rabin8本身是如何在细胞中被激活的尚不清楚。最近，Rabin8被发现是Rab11的下游效应物，它控制着囊泡从循环内体中退出。以gtp结合形式存在的Rab11可以刺激Rabin8的GEF活性。因此，Rab11-Rabin8-Rab8相互作用将囊泡从供体室产生到传递到质膜。在这里，我们描述了我们用来表达和纯化几种rabb蛋白的方法，并测定了Rab11对Rabin8 GEF活性的动力学激活的影响。
The Rab family of small GTPases acts as molecular switches that control various stages of vesicular transport. Rab8 functions in exocytic trafficking from thetrans-Golgi network (TGN) and recycling endosomes to the plasma membrane. Rabin8 is a major guanine nucleotide exchange factor (GEF) for Rab8. It activates Rab8 by catalyzing its GDP release for subsequent GTP loading. However, how Rabin8 itself is activated in cells is unclear. Recently, it was found that Rabin8 is a downstream effector of Rab11, which controls vesicle exit from the recycling endosomes. Rab11, in its GTP-bound form, stimulates the GEF activity of Rabin8. The Rab11–Rabin8–Rab8 interactions thus couple vesicle generation from the donor compartment to its delivery to plasma membrane. Here we describe the methods we used to express and purify several Rab proteins, and to assay for the effect of Rab11 in the kinetic activation of Rabin8 GEF activity.