Dynamics of the G Protein-coupled Vasopressin V2 Receptor Signaling Network Revealed by Quantitative Phosphoproteomics
Dynamics of the G Protein-coupled Vasopressin V2 Receptor Signaling Network Revealed by Quantitative Phosphoproteomics
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DOI：
10.1074/mcp.m111.014613
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发表时间：
2012-02-01
影响因子：
7
通讯作者：
Knepper, Mark A.
中科院分区：
文献类型：
--
作者：
Hoffert, Jason D.;Pisitkun, Trairak;Knepper, Mark A.
G蛋白偶联受体（GPCR）调节多种生理过程，并且许多人类疾病是由于GPCR信号传导的缺陷。为了鉴定原型G（s）偶联GPCR（加压素V2受体）下游信号网络的动态响应，我们在大鼠肾脏分离细胞中暴露于生理浓度的加压素后的四个时间点进行了多重复、定量磷酸化蛋白质组学和iTRAQ标记。从2，783种蛋白质中鉴定出12，167种磷酸肽，其中273种在加压素的丰度上发生了显著变化。磷酸肽时间进程和基因本体论术语的二维聚类显示，配体与V2受体的结合不仅仅影响生理条件下的典型环磷酸腺苷-蛋白激酶A和抑制蛋白途径。调控蛋白包括肌动蛋白细胞骨架重塑、细胞间粘附、丝裂原活化蛋白激酶信号传导、Wnt/β-连环蛋白信号传导和凋亡途径的关键组分。这些数据表明，加压素可以调节一系列的细胞功能远远超出其经典的作用，在调节水和溶质的运输。这些结果极大地扩展了GPCR信号在生理背景下的当前观点，并为这种信号网络在多囊肾疾病等疾病中的潜在作用提供了新的线索。最后，我们提供了一个动态定量数据的生理调节磷酸化位点的在线资源（http：//helixweb.nih.gov/ESBL/Database/TiPD/index.html）。Molecular & Cellular Proteomics 11：10.1074/mcp. M111.014613，1-15，2012.
G protein-coupled receptors (GPCRs) regulate diverse physiological processes, and many human diseases are due to defects in GPCR signaling. To identify the dynamic response of a signaling network downstream from a prototypical G(s)-coupled GPCR, the vasopressin V2 receptor, we have carried out multireplicate, quantitative phosphoproteomics with iTRAQ labeling at four time points following vasopressin exposure at a physiological concentration in cells isolated from rat kidney. A total of 12,167 phosphopeptides were identified from 2,783 proteins, with 273 changing significantly in abundance with vasopressin. Two-dimensional clustering of phosphopeptide time courses and Gene Ontology terms revealed that ligand binding to the V2 receptor affects more than simply the canonical cyclic adenosine monophosphate-protein kinase A and arrestin pathways under physiological conditions. The regulated proteins included key components of actin cytoskeleton remodeling, cell-cell adhesion, mitogen-activated protein kinase signaling, Wnt/beta-catenin signaling, and apoptosis pathways. These data suggest that vasopressin can regulate an array of cellular functions well beyond its classical role in regulating water and solute transport. These results greatly expand the current view of GPCR signaling in a physiological context and shed new light on potential roles for this signaling network in disorders such as polycystic kidney disease. Finally, we provide an online resource of physiologically regulated phosphorylation sites with dynamic quantitative data (http://helixweb.nih.gov/ESBL/Database/TiPD/index.html). Molecular & Cellular Proteomics 11: 10.1074/mcp.M111.014613, 1-15, 2012.