Role of Adaptor Proteins and Clathrin in the Trafficking of Human Kidney Anion Exchanger 1 (kAE1) to the Cell Surface
Role of Adaptor Proteins and Clathrin in the Trafficking of Human Kidney Anion Exchanger 1 (kAE1) to the Cell Surface
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接头蛋白和网格蛋白在人肾阴离子交换器 1 (kAE1) 运输至细胞表面中的作用
DOI：
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发表时间：
2014
期刊：
影响因子：
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通讯作者：
P. Yenchitsomanus
中科院分区：
文献类型：
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作者：
Mutita Junking;N. Sawasdee;N. Duangtum;B. Cheunsuchon;T. Limjindaporn;P. Yenchitsomanus
肾脏阴离子交换器 1 (kAE1) 通过介导远端肾单位 α-嵌入细胞基底外侧膜上的氯离子/碳酸氢根 (Cl−/HCO3−) 交换，在酸碱稳态中发挥重要作用。由编码 kAE1 的 SLC4A1 突变引起的 kAE1 细胞内运输受损会导致肾脏疾病——远端肾小管酸中毒 (dRTA)。然而，目前尚不清楚 kAE1 从跨高尔基体网络 (TGN) 到基底外侧膜的细胞内分选和运输是如何发生的。在这里，我们研究了基底外侧相关分选蛋白（包括接头蛋白 (AP) 复合物的 mu1 亚基、网格蛋白和蛋白激酶 D）对极化和非极化肾细胞中 kAE1 运输的作用。通过使用 RNA 干扰、免疫共沉淀、基于黄色荧光蛋白的蛋白片段互补测定和免疫荧光染色，我们证明 AP-1 mu1A、AP-3 mu1、AP-4 mu1 和网格蛋白（但不是 AP-1 mu1B、PKD1 或 PKD2）在 kAE1 的细胞内分选和运输中发挥关键作用。我们还通过双重免疫荧光染色证明了 kAE1 和基底外侧相关分选蛋白在人肾组织中的共定位。这些发现表明，AP-1 mu1A、AP-3 mu1、AP-4 mu1 和网格蛋白是 kAE1 分选和从 TGN 运输到分泌酸的 α-嵌入细胞的基底外侧膜所必需的。
Kidney anion exchanger 1 (kAE1) plays an important role in acid–base homeostasis by mediating chloride/bicarbornate (Cl−/HCO3−) exchange at the basolateral membrane of α‐intercalated cells in the distal nephron. Impaired intracellular trafficking of kAE1 caused by mutations of SLC4A1 encoding kAE1 results in kidney disease – distal renal tubular acidosis (dRTA). However, it is not known how the intracellular sorting and trafficking of kAE1 from trans‐Golgi network (TGN) to the basolateral membrane occurs. Here, we studied the role of basolateral‐related sorting proteins, including the mu1 subunit of adaptor protein (AP) complexes, clathrin and protein kinase D, on kAE1 trafficking in polarized and non‐polarized kidney cells. By using RNA interference, co‐immunoprecipitation, yellow fluorescent protein‐based protein fragment complementation assays and immunofluorescence staining, we demonstrated that AP‐1 mu1A, AP‐3 mu1, AP‐4 mu1 and clathrin (but not AP‐1 mu1B, PKD1 or PKD2) play crucial roles in intracellular sorting and trafficking of kAE1. We also demonstrated colocalization of kAE1 and basolateral‐related sorting proteins in human kidney tissues by double immunofluorescence staining. These findings indicate that AP‐1 mu1A, AP‐3 mu1, AP‐4 mu1 and clathrin are required for kAE1 sorting and trafficking from TGN to the basolateral membrane of acid‐secreting α‐intercalated cells.
影响因子：
11.8
作者：
Gravotta, Diego;Carvajal-Gonzalez, Jose Maria;Mattera, Rafael;Deborde, Sylvie;Banfelder, Jason R.;Bonifacino, Juan S.;Rodriguez-Boulan, Enrique
通讯作者：
Rodriguez-Boulan, Enrique