Identification of Rab6 as an N-ethylmaleimide-sensitive fusion protein-binding protein

被引:22
作者
Han, SY
Park, DY
Park, SD
Hong, SH [1 ]
机构
[1] Seoul Natl Univ, Inst Mol Biol & Genet, Seoul 151742, South Korea
[2] Seoul Natl Univ, Sch Biol Sci, Seoul 151742, South Korea
关键词
NSF; Rab protein; small G-protein; vesicular transport;
D O I
10.1042/0264-6021:3520165
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In this study we show the interaction of N-ethylmaleimide-sensitive fusion protein (NSF) with a small GT:P-binding protein, Rab6. NSF is an ATPase involved in the vesicular transport within eukaryotic cells. Using the yeast two-hybrid system, we have isolated new NSF-binding proteins from the rat lung cDNA library. One of them was Rab6, which is invoked in the vesicular transport within the Golgi and trans-Golgi network as a Ras-like GTPase, We demonstrated that the N-terminal domain of NSF interacted with the C-terminal domain of Rab6, and these proteins were co-immunoprecipitated from the rat brain extract. This interaction was maintained preferentially in the presence of hydrolysable ATP. Recombinant NSF-His(6) can also bind to C-terminal Rab6-glutathione S-transferase under the conditions to allow the ATP hydrolysis. Surprisingly, Rab6 stimulates the ATPase activity of NSF by approx. 2-fold as does alpha -soluble NSF attachment protein receptor. Anti-Rab6 polyclonal antibodies significantly inhibited the Rab6-stimulated ATPase activity of NSF. Furthermore, we found that Rab3 and Rab4 can also associate with NSF and stimulate its ATPase activity. Taken together, we propose a model in which Rab can form an ATP hydrolysis-regulated complex with NSF, and function as a signalling molecule to deliver the signal of vesicle fusion through the interaction with NSF.
引用
收藏
页码:165 / 173
页数:9
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