Large-scale proteomics and phosphoproteomics of urinary exosomes.

Published

Journal Article

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/.

Full Text

Duke Authors

Cited Authors

  • Gonzales, PA; Pisitkun, T; Hoffert, JD; Tchapyjnikov, D; Star, RA; Kleta, R; Wang, NS; Knepper, MA

Published Date

  • February 2009

Published In

Volume / Issue

  • 20 / 2

Start / End Page

  • 363 - 379

PubMed ID

  • 19056867

Pubmed Central ID

  • 19056867

Electronic International Standard Serial Number (EISSN)

  • 1533-3450

Digital Object Identifier (DOI)

  • 10.1681/ASN.2008040406

Language

  • eng

Conference Location

  • United States