Mapping the H(+) (V)-ATPase interactome: identification of proteins involved in trafficking, folding, assembly and phosphorylation.
Mapping the H(+) (V)-ATPase interactome: identification of proteins involved in trafficking, folding, assembly and phosphorylation.
复制标题
DOI：
10.1038/srep14827
复制
发表时间：
2015-10-07
影响因子：
4.6
通讯作者：
Brown D
中科院分区：
文献类型：
--
作者：
Merkulova M;Păunescu TG;Azroyan A;Marshansky V;Breton S;Brown D
V-ATP 酶（H+ ATP 酶）是多亚基、ATP 依赖性质子泵，可调节几乎所有真核生物中的 pH 稳态。它们参与关键的细胞生物过程，包括囊泡运输、内体 pH 传感、膜融合和细胞内信号传导。它们还在肾酸排泄和血液 pH 平衡、男性生育能力、骨重塑、突触传递、嗅觉和听力方面发挥着关键的系统作用。此外，V-ATP酶功能障碍会导致或加重各种其他疾病，但人们对调节这些不同V-ATP酶功能的复杂蛋白质相互作用知之甚少。因此，我们进行了蛋白质组分析来鉴定 V-ATP 酶相关蛋白并构建 V-ATP 酶相互作用组。我们对肾组织的分析揭示了 V-ATP 酶相关蛋白簇参与蛋白质质量控制、复杂组装和细胞内运输。本研究首次发现 ARHGEF7、DMXL1、EZR、NCOA7、OXR1、RPS6KA3、SNX27 和含有 TCP1 复合物 (CCT) 的伴侣蛋白的 9 个亚基与 V-ATPase 相互作用。两种相互作用蛋白 DMXL1 和 WDR7 的敲低抑制了肾细胞系中 V-ATP 酶介导的细胞内囊泡酸化，这为我们的相互作用组作为功能重要的新型 V-ATP 酶调节蛋白的筛选的效用提供了验证。因此，我们的数据为 V-ATP 酶细胞生物学和（病理）生理学分析提供了新的见解和方向。
V-ATPases (H+ ATPases) are multisubunit, ATP-dependent proton pumps that regulate pH homeostasis in virtually all eukaryotes. They are involved in key cell biological processes including vesicle trafficking, endosomal pH sensing, membrane fusion and intracellular signaling. They also have critical systemic roles in renal acid excretion and blood pH balance, male fertility, bone remodeling, synaptic transmission, olfaction and hearing. Furthermore, V-ATPase dysfunction either results in or aggravates various other diseases, but little is known about the complex protein interactions that regulate these varied V-ATPase functions. Therefore, we performed a proteomic analysis to identify V-ATPase associated proteins and construct a V-ATPase interactome. Our analysis using kidney tissue revealed V-ATPase-associated protein clusters involved in protein quality control, complex assembly and intracellular trafficking. ARHGEF7, DMXL1, EZR, NCOA7, OXR1, RPS6KA3, SNX27 and 9 subunits of the chaperonin containing TCP1 complex (CCT) were found to interact with V-ATPase for the first time in this study. Knockdown of two interacting proteins, DMXL1 and WDR7, inhibited V-ATPase-mediated intracellular vesicle acidification in a kidney cell line, providing validation for the utility of our interactome as a screen for functionally important novel V-ATPase-regulating proteins. Our data, therefore, provide new insights and directions for the analysis of V-ATPase cell biology and (patho)physiology.