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Large-scale proteomics and phosphoproteomics of urinary exosomes.

Publication ,  Journal Article
Gonzales, PA; Pisitkun, T; Hoffert, JD; Tchapyjnikov, D; Star, RA; Kleta, R; Wang, NS; Knepper, MA
Published in: J Am Soc Nephrol
February 2009

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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Published In

J Am Soc Nephrol

DOI

EISSN

1533-3450

Publication Date

February 2009

Volume

20

Issue

2

Start / End Page

363 / 379

Location

United States

Related Subject Headings

  • Urology & Nephrology
  • Urine
  • Thiazides
  • Proteomics
  • Proteome
  • Phosphorylation
  • Phosphoproteins
  • Mass Spectrometry
  • Male
  • Humans
 

Citation

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Gonzales, P. A., Pisitkun, T., Hoffert, J. D., Tchapyjnikov, D., Star, R. A., Kleta, R., … Knepper, M. A. (2009). Large-scale proteomics and phosphoproteomics of urinary exosomes. J Am Soc Nephrol, 20(2), 363–379. https://doi.org/10.1681/ASN.2008040406
Gonzales, Patricia A., Trairak Pisitkun, Jason D. Hoffert, Dmitry Tchapyjnikov, Robert A. Star, Robert Kleta, Nam Sun Wang, and Mark A. Knepper. “Large-scale proteomics and phosphoproteomics of urinary exosomes.J Am Soc Nephrol 20, no. 2 (February 2009): 363–79. https://doi.org/10.1681/ASN.2008040406.
Gonzales PA, Pisitkun T, Hoffert JD, Tchapyjnikov D, Star RA, Kleta R, et al. Large-scale proteomics and phosphoproteomics of urinary exosomes. J Am Soc Nephrol. 2009 Feb;20(2):363–79.
Gonzales, Patricia A., et al. “Large-scale proteomics and phosphoproteomics of urinary exosomes.J Am Soc Nephrol, vol. 20, no. 2, Feb. 2009, pp. 363–79. Pubmed, doi:10.1681/ASN.2008040406.
Gonzales PA, Pisitkun T, Hoffert JD, Tchapyjnikov D, Star RA, Kleta R, Wang NS, Knepper MA. Large-scale proteomics and phosphoproteomics of urinary exosomes. J Am Soc Nephrol. 2009 Feb;20(2):363–379.

Published In

J Am Soc Nephrol

DOI

EISSN

1533-3450

Publication Date

February 2009

Volume

20

Issue

2

Start / End Page

363 / 379

Location

United States

Related Subject Headings

  • Urology & Nephrology
  • Urine
  • Thiazides
  • Proteomics
  • Proteome
  • Phosphorylation
  • Phosphoproteins
  • Mass Spectrometry
  • Male
  • Humans