Journal of the American Society of Nephrology
2008 JASN IMPACT FACTOR 7.505 HOME   AUTHOR INFO   EDITORIAL BOARD   SUBSCRIBE   FEEDBACK   ALERTS   HELP 
    advanced
CURRENT ISSUE ARCHIVES JASN Express ONLINE SUBMISSION


Published ahead of print on December 3, 2008
J Am Soc Nephrol 20: 363-379, 2009
© 2009 American Society of Nephrology
doi: 10.1681/ASN.2008040406
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental Data
Right arrow All Versions of this Article:
ASN.2008040406v1
20/2/363    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Gonzales, P. A.
Right arrow Articles by Knepper, M. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Gonzales, P. A.
Right arrow Articles by Knepper, M. A.

BASIC RESEARCH

Large-Scale Proteomics and Phosphoproteomics of Urinary Exosomes

Patricia A. Gonzales*,{dagger}, Trairak Pisitkun*, Jason D. Hoffert*, Dmitry Tchapyjnikov*, Robert A. Star{ddagger}, Robert Kleta§,||,¶, Nam Sun Wang{dagger} and Mark A. Knepper*

* Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung, and Blood Institute, {ddagger} Renal Diagnostics and Therapeutics Unit, National Institute of Diabetes and Digestive and Kidney Diseases, § Section of Human Biochemical Genetics, Medical Genetics Branch, National Human Genome Research Institute, and || Office of Rare Diseases, Office of the Director, National Institutes of Health, Bethesda, and {dagger} Department of Chemical and Biomolecular Engineering, University of Maryland, College Park, Maryland; and London Epithelial Group, Centre for Nephrology, University College London, London, United Kingdom

Correspondence: Dr. Mark A. Knepper, 10 Center Drive, MSC-1603, National Institutes of Health, Bethesda, MD 20892-1603. Phone: 301-496-3064; Fax: 301-402-1443; E-mail: knep{at}helix.nih.gov

Received for publication April 21, 2008. Accepted for publication July 30, 2008.

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/.




This article has been cited by other articles:


Home page
 Hum Mol GenetHome page
A. Kurbegovic, O. Cote, M. Couillard, C. J. Ward, P. C. Harris, and M. Trudel
Pkd1 transgenic mice: adult model of polycystic kidney disease with extrarenal and renal phenotypes
Hum. Mol. Genet., January 19, 2010; (2010) ddp588v2.
[Abstract] [Full Text] [PDF]

Home page
 Nephrol Dial TransplantHome page
V. Thongboonkerd
Current status of renal and urinary proteomics: ready for routine clinical application?
Nephrol. Dial. Transplant., January 1, 2010; 25(1): 11 - 16.
[Full Text] [PDF]

Home page
 Am. J. Physiol. Renal Physiol.Home page
H. Sonoda, N. Yokota-Ikeda, S. Oshikawa, Y. Kanno, K. Yoshinaga, K. Uchida, Y. Ueda, K. Kimiya, S. Uezono, A. Ueda, et al.
Decreased abundance of urinary exosomal aquaporin-1 in renal ischemia-reperfusion injury
Am J Physiol Renal Physiol, October 1, 2009; 297(4): F1006 - F1016.
[Abstract] [Full Text] [PDF]

Home page
 J. Am. Soc. Nephrol.Home page
M. A. Knepper
Common Sense Approaches to Urinary Biomarker Study Design
J. Am. Soc. Nephrol., June 1, 2009; 20(6): 1175 - 1178.
[Full Text] [PDF]


HOME CURRENT ISSUE ARCHIVES JASN Express ONLINE SUBMISSION AUTHOR INFO
EDITORIAL BOARD SUBSCRIBE FEEDBACK ALERTS HELP

Copyright © 2009 by the American Society of Nephrology. Online ISSN: 1533-3450 Print ISSN: 1046-6673