Retromer- and WASH-dependent sorting of nutrient transporters requires a multivalent interaction network with ANKRD50
Retromer- and WASH-dependent sorting of nutrient transporters requires a multivalent interaction network with ANKRD50
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DOI：
10.1242/jcs.196758
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发表时间：
2017-01-15
影响因子：
4
通讯作者：
Steinberg, Florian
中科院分区：
文献类型：
--
作者：
Kvainickas, Arunas;Orgaz, Ana Jimenez;Steinberg, Florian
逆转录酶和相关的肌动蛋白聚合 WASH 复合物对于多种整合膜蛋白的内吞再循环至关重要。逆转录体亚基 VPS35 中的遗传性帕金森病致病点突变 (D620N) 扰乱逆转录体与 WASH 复合体以及未表征的含锚蛋白重复结构域的蛋白 50 (ANKRD50) 的关联。在这里，我们坚定地将 ANKRD50 确立为 SNX27-retromer-WASH 超级复合物的新的重要组成部分。 HeLa 或 U2OS 细胞中 ANKRD50 的耗竭反映了在抑制 SNX27、逆转录体或 WASH 复合物成分后观察到的多种跨膜蛋白的内体到细胞表面再循环的丧失。对 ANKRD50 耗尽细胞的细胞表面蛋白质组进行基于质谱的定量，鉴定出 SLC1A 家族的氨基酸转运蛋白（其中包括 SLC1A4）作为依赖于 ANKRD50 和逆转录酶进行内吞回收的额外货物分子。从机制上讲，我们表明 ANKRD50 同时参与 SNX27-retromer-WASH 复杂机制的多个部分，形成一个直接且协作的相互作用网络，该网络是有效回收营养转运蛋白 GLUT1（也称为 SLC2A1）和 SLC1A4 以及潜在的许多其他表面蛋白所必需的。
Retromer and the associated actin-polymerizing WASH complex are essential for the endocytic recycling of a wide range of integral membrane proteins. A hereditary Parkinson's-disease-causing point mutation (D620N) in the retromer subunit VPS35 perturbs retromer's association with the WASH complex and also with the uncharacterized protein ankyrin-repeat-domain-containing protein 50 (ANKRD50). Here, we firmly establish ANKRD50 as a new and essential component of the SNX27-retromer-WASH super complex. Depletion of ANKRD50 in HeLa or U2OS cells phenocopied the loss of endosome-to-cell-surface recycling of multiple transmembrane proteins seen upon suppression of SNX27, retromer or WASH-complex components. Mass-spectrometry-based quantification of the cell surface proteome of ANKRD50-depleted cells identified amino acid transporters of the SLC1A family, among them SLC1A4, as additional cargo molecules that depend on ANKRD50 and retromer for their endocytic recycling. Mechanistically, we show that ANKRD50 simultaneously engages multiple parts of the SNX27-retromer- WASH complex machinery in a direct and co-operative interaction network that is needed to efficiently recycle the nutrient transporters GLUT1 (also known as SLC2A1) and SLC1A4, and potentially many other surface proteins.