Rab11 family interacting protein 2 associates with myosin Vb and regulates plasma membrane recycling
Rab11 family interacting protein 2 associates with myosin Vb and regulates plasma membrane recycling
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DOI：
10.1074/jbc.m209270200
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发表时间：
2002-12-27
影响因子：
4.8
通讯作者：
Goldenring, JR
中科院分区：
文献类型：
--
作者：
Hales, CM;Vaerman, JP;Goldenring, JR
质膜循环是维持细胞膜组成的一个重要过程。运动蛋白Myosin VB通过与Rab11a结合来调节质膜循环。肌球蛋白VB尾部的过表达扰乱了质膜循环系统的运输，导致Rab11a在极化和非极化细胞中的积累。我们研究了Rab11家族相互作用蛋白2(Rab11-FIP2)与作为Rab11a和myosin VB之间的接头蛋白的肌球蛋白VB之间的关系。免疫荧光研究表明，内源性Rab11-FIP2与绿色荧光蛋白-肌球蛋白VB Tail在Madin-Darby犬肾(MDCK)细胞中过表达。酵母双杂交实验表明，Rab11-FIP2的129-356位氨基酸是与肌球蛋白VB尾结合的重要氨基酸。体外结合实验和在MDCK和HeLa细胞中的共转染实验证实了这一结果，但进一步提纯了Rab11-FIP2与129-290氨基酸的结合部位。与肌球蛋白VB一样，功能研究表明Rab11-FIP2对正常的质膜循环也是重要的。绿色荧光蛋白Rab11-FIP2(129-512)缺乏其氨基末端的C2结构域，其功能是一个显性的负作用截短，导致Rab11a的积聚，并扰乱MDCK细胞的IgA转运和HeLa细胞的转铁蛋白转运。肌球蛋白VB和Rab11-FIP2与Rab11a的三元结合表明，一个多聚体蛋白复合体参与了囊泡通过质膜循环系统的运输。
Plasma membrane recycling is an important process necessary for maintaining membrane composition. The motor protein myosin Vb regulates plasma membrane recycling through its association with Rab11a. Overexpression of the tail of myosin Vb disrupts trafficking out of plasma membrane recycling systems and leads to the accumulation of Rab11a in both polarized and non-polarized cells. We have investigated the association of Rab11 family interacting protein 2 (Rab11-FIP2) with myosin Vb as an adapter protein between Rab11a and myosin Vb. Immunofluorescence studies indicated a colocalization of endogenous Rab11-FIP2 with green fluorescent protein-myosin Vb tail overexpressed in Madin-Darby canine kidney (MDCK) cells. Yeast two hybrid assays showed that amino acids 129-356 of Rab11-FIP2 were important for binding to myosin Vb tail. In vitro association assays and co-transfection experiments in both MDCK and HeLa cells confirmed this result but further refined the binding site to amino acids 129-290 of Rab11-FIP2. Like myosin Vb, functional studies indicated that Rab11-FIP2 is also important for normal plasma membrane recycling. Green fluorescent protein-Rab11-FIP2 (129-512), which lacks its amino-terminal C2 domain, functioned as a dominant negative acting truncation that caused accumulation of Rab11a and disrupted IgA trafficking in MDCK cells and transferrin trafficking in HeLa cells. The ternary association of myosin Vb and Rab11-FIP2 with Rab11a suggests that a multimeric protein complex is involved in vesicle trafficking through plasma membrane recycling systems.