Identification of UT-A1- and AQP2-interacting proteins in rat inner medullary collecting duct

Chou, Chung‐Lin; Hwang, Gloria Yu-yan; Hageman, Daniel J.; Han, L.; Agrawal, Prashasti; Pisitkun, Trairak; Knepper, Mark A.

doi:10.1152/ajpcell.00082.2017

Cited by 17 publications

(17 citation statements)

References 67 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Immunoprecipitated proteins were excised from SDS-PAGE gels and digested as previously described (Chou et al, 2018 ). Briefly, the excised gel pieces were washed twice in 50% acetonitrile (ACN) for 30 min and 100% ACN for 30 min.…”

Section: Methodsmentioning

confidence: 99%

THBS1/CD47 Modulates the Interaction of γ-Catenin With E-Cadherin and Participates in Epithelial–Mesenchymal Transformation in Lipid Nephrotoxicity

Gao

Yang

Zhang

et al. 2021

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

Hyperlipidemia, an important risk factor for cardiovascular and end-stage renal diseases, often aggravates renal injury and compromises kidney function. Here, histological analysis of human kidney samples revealed that high lipid levels induced the development of renal fibrosis. To elucidate the mechanism underlying lipid nephrotoxicity, we used two types of mouse models (Apoe−/− and C57BL/6 mice fed a 45 and 60% high-fat diet, respectively). Histological analysis of kidney tissues revealed high-lipid-induced renal fibrosis and inflammation; this was confirmed by examining fibrotic and inflammatory marker expression using Western blotting and real-time polymerase chain reaction. Oxidized low-density lipoprotein (OX-LDL) significantly induced the fibrotic response in HK-2 tubular epithelial cells. RNA-sequencing and Gene Ontology analysis of differentially expressed mRNAs in OX-LDL-treated HK-2 tubular epithelial cells and real-time PCR validation in Apoe−/− mice showed that the expression of thrombospondin-1 (THBS1) in the high-fat group was significantly higher than that of the other top known genes, along with significant overexpression of its receptor CD47. THBS1 knockdown cells verified its relation to OX-LDL-induced fibrosis and inflammation. Liquid chromatography tandem mass spectrometry and STRING functional protein association network analyses predicted that THBS1/CD47 modulated the interaction between γ-catenin and E-cadherin and was involved in epithelial–mesenchymal transition, which was supported by immunoprecipitation and immunohistochemistry. CD47 downregulation following transfection with small-hairpin RNA in OX-LDL-treated tubular epithelial cells and treatment with anti-CD47 antibody restored the expression of E-cadherin and attenuated renal injury, fibrosis, and inflammatory response in OX-LDL-treated cells and in type 2 diabetes mellitus. These findings indicate that CD47 may serve as a potential therapeutic target in long-term lipid-induced kidney injury.

show abstract

Section: Methodsmentioning

confidence: 99%

THBS1/CD47 Modulates the Interaction of γ-Catenin With E-Cadherin and Participates in Epithelial–Mesenchymal Transformation in Lipid Nephrotoxicity

Gao

Yang

Zhang

et al. 2021

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

show abstract

“…Proteins that form complexes with AQP2 are likely to be phosphorylated by the same protein kinases as AQP2. Consequently, we asked what proteins identified as AQP2-binding proteins in a previous mass spectrometry-based study (15) have sites that showed increases in phosphorylation. These proteins are shown in Table 6, with the protein kinases that phosphorylate them.…”

Section: F796 Vasopressin Signaling In the Collecting Ductmentioning

confidence: 99%

Phosphoproteomic identification of vasopressin V2 receptor-dependent signaling in the renal collecting duct

Deshpande

Kao

Raghuram

et al. 2019

American Journal of Physiology-Renal Physiology

Self Cite

View full text Add to dashboard Cite

Vasopressin controls water balance largely through PKA-dependent effects to regulate the collecting duct water channel aquaporin-2 (AQP2). Although considerable information has accrued regarding the regulation of water and solute transport in collecting duct cells, information is sparse regarding the signaling connections between PKA and transport responses. Here, we exploited recent advancements in protein mass spectrometry to perform a comprehensive, multiple-replicate analysis of changes in the phosphoproteome of native rat inner medullary collecting duct cells in response to the vasopressin V2 receptor-selective agonist 1-desamino-8D-arginine vasopressin. Of the 10,738 phosphopeptides quantified, only 156 phosphopeptides were significantly increased in abundance, and only 63 phosphopeptides were decreased, indicative of a highly selective response to vasopressin. The list of upregulated phosphosites showed several general characteristics: 1) a preponderance of sites with basic (positively charged) amino acids arginine (R) and lysine (K) in position −2 and −3 relative to the phosphorylated amino acid, consistent with phosphorylation by PKA and/or other basophilic kinases; 2) a greater-than-random likelihood of sites previously demonstrated to be phosphorylated by PKA; 3) a preponderance of sites in membrane proteins, consistent with regulation by membrane association; and 4) a greater-than-random likelihood of sites in proteins with class I COOH-terminal PDZ ligand motifs. The list of downregulated phosphosites showed a preponderance of those with proline in position +1 relative to the phosphorylated amino acid, consistent with either downregulation of proline-directed kinases (e.g., MAPKs or cyclin-dependent kinases) or upregulation of one or more protein phosphatases that selectively dephosphorylate such sites (e.g., protein phosphatase 2A). The phosphoproteomic data were used to create a web resource for the investigation of G protein-coupled receptor signaling and regulation of AQP2-mediated water transport.

show abstract

“…The effect of vasopressin on protein-protein interactions in the collecting duct was also recently studied. By focusing on the UT-A1, a known urea transporter regulated by vasopressin, Chou et al (30) used chemical cross-linking, followed by immunoprecipitation of UT-A1 in native rat IMCD cells, incubated with or without dDAVP for 15 min to study its interacting partner proteins. LC-MS/MS analysis revealed 25 partner proteins significantly changed in its binding with UT-A1 in response to dDAVP.…”

Section: Iii) S264mentioning

confidence: 99%

From Molecules to Mechanisms: Functional Proteomics and Its Application to Renal Tubule Physiology

Rinschen

Limbutara

Knepper

et al. 2018

Physiological Reviews

Self Cite

View full text Add to dashboard Cite

Classical physiological studies using electrophysiological, biophysical, biochemical, and molecular techniques have created a detailed picture of molecular transport, bioenergetics, contractility and movement, and growth, as well as the regulation of these processes by external stimuli in cells and organisms. Newer systems biology approaches are beginning to provide deeper and broader understanding of these complex biological processes and their dynamic responses to a variety of environmental cues. In the past decade, advances in mass spectrometry-based proteomic technologies have provided invaluable tools to further elucidate these complex cellular processes, thereby confirming, complementing, and advancing common views of physiology. As one notable example, the application of proteomics to study the regulation of kidney function has yielded novel insights into the chemical and physical processes that tightly control body fluids, electrolytes, and metabolites to provide optimal microenvironments for various cellular and organ functions. Here, we systematically review, summarize, and discuss the most significant key findings from functional proteomic studies in renal epithelial physiology. We also identify further improvements in technological and bioinformatics methods that will be essential to advance precision medicine in nephrology.

show abstract

Identification of UT-A1- and AQP2-interacting proteins in rat inner medullary collecting duct

Cited by 17 publications

References 67 publications

THBS1/CD47 Modulates the Interaction of γ-Catenin With E-Cadherin and Participates in Epithelial–Mesenchymal Transformation in Lipid Nephrotoxicity

THBS1/CD47 Modulates the Interaction of γ-Catenin With E-Cadherin and Participates in Epithelial–Mesenchymal Transformation in Lipid Nephrotoxicity

Phosphoproteomic identification of vasopressin V2 receptor-dependent signaling in the renal collecting duct

From Molecules to Mechanisms: Functional Proteomics and Its Application to Renal Tubule Physiology

Contact Info

Product

Resources

About