Large-scale Proteomics and Phosphoproteomics of Urinary Exosomes

Overview

Journal J Am Soc Nephrol

Specialty Nephrology

Date 2008 Dec 6

PMID 19056867

Citations 368

Authors

Patricia A Gonzales

Trairak Pisitkun

Jason D Hoffert

Dmitry Tchapyjnikov

Robert A Star

Robert Kleta

Nam Sun Wang

Mark A Knepper

Affiliations

Soon will be listed here.

Abstract

Normal human urine contains large numbers of exosomes, which are 40- to 100-nm vesicles that originate as the internal vesicles in multivesicular bodies from every renal epithelial cell type facing the urinary space. Here, we used LC-MS/MS to profile the proteome of human urinary exosomes. Overall, the analysis identified 1132 proteins unambiguously, including 177 that are represented on the Online Mendelian Inheritance in Man database of disease-related genes, suggesting that exosome analysis is a potential approach to discover urinary biomarkers. We extended the proteomic analysis to phosphoproteomic profiling using neutral loss scanning, and this yielded multiple novel phosphorylation sites, including serine-811 in the thiazide-sensitive Na-Cl co-transporter, NCC. To demonstrate the potential use of exosome analysis to identify a genetic renal disease, we carried out immunoblotting of exosomes from urine samples of patients with a clinical diagnosis of Bartter syndrome type I, showing an absence of the sodium-potassium-chloride co-transporter 2, NKCC2. The proteomic data are publicly accessible at http://dir.nhlbi.nih.gov/papers/lkem/exosome/.

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