Identification of the functional domains of yeast sorting nexins Vps5p and Vps17p.
Identification of the functional domains of yeast sorting nexins Vps5p and Vps17p.
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DOI：
10.1091/mbc.02-05-0064
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发表时间：
2002-08
影响因子：
3.3
通讯作者：
M. Seaman;Hazel P. Williams
中科院分区：
文献类型：
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作者：
M. Seaman;Hazel P. Williams
分选连接蛋白 (Snxs) 是最近发现的一个保守的亲水性细胞质蛋白家族，已发现它们与内吞系统的膜相关，并且参与许多内体膜蛋白的运输，包括表皮生长因子受体和转铁蛋白受体。 Snx 蛋白的部分定义是 p40 phox 同源结构域的存在，该结构域最近被证明可以结合磷脂酰肌醇 3-磷酸。大多数 Snx 蛋白在其羧基末端部分还包含一个预测的卷曲螺旋结构域，并且已被证明可以与 Snx 家族的其他成员形成二聚体。酵母分选连接蛋白 Vps5p 和 Vps17p 形成二聚体，也是介导羧肽酶 Y 受体 Vps10p 的内体到高尔基体转运的逆转录复合体的组成部分。为了在功能上定义酵母分选连接蛋白 Vps5p 和 Vps17p 的不同结构域，我们生成了各种截断来检查 Vps5p/Vps17p 的不同结构域在各自功能中发挥的作用。在此，我们证明 Vps5p 和 Vps17p 的 C 端部分（包含卷曲螺旋结构域）对于它们的相互作用是必要且充分的。我们还将逆转录聚合体组装结构域映射到 Vps5p 的 N 端一半，并发现 Vps17p 与 Vps5p 的结合可协同 Vps5p 和其他逆转录聚合体组件之间的相互作用。此外，我们还检查了 Vps5p 的哪些结构域对于膜关联是必需的。
Sorting nexins (Snxs) are a recently discovered family of conserved hydrophilic cytoplasmic proteins that have been found associated with membranes of the endocytic system and that are implicated in the trafficking of many endosomal membrane proteins, including the epidermal growth factor receptor and transferrin receptor. Snx proteins are partly defined by the presence of a p40 phox homology domain that has recently been shown to bind phosphatidylinositol 3-phosphate. Most Snx proteins also contain a predicted coiled-coils domain in the carboxyl-terminal half of the protein and have been shown to form dimers with other members of the Snx family. The yeast sorting nexins Vps5p and Vps17p form a dimer and are also components of the retromer complex that mediates endosome-to-Golgi transport of the carboxypeptidase Y receptor Vps10p. To functionally define the different domains of the yeast sorting nexins Vps5p and Vps17p, we have generated various truncations to examine the role that the different domains of Vps5p/Vps17p play in their respective functions. Herein, we show that the C-terminal halves of Vps5p and Vps17p, which contain the coiled-coils domains, are necessary and sufficient for their interaction. We have also mapped the retromer assembly domain to the N-terminal half of Vps5p and found that binding of Vps5p by Vps17p synergizes the interaction between Vps5p and other retromer components. Additionally, we have examined which domain(s) of Vps5p is necessary for membrane association.