Document Detail

Proteomic analysis of brush-border membrane vesicles isolated from purified proximal convoluted tubules.
MedLine Citation:
PMID:  20219825     Owner:  NLM     Status:  MEDLINE    
The renal proximal convoluted tubule is the primary site of water, electrolyte and nutrient reabsorption and of active secretion of selected molecules. Proteins in the apical brush-border membrane facilitate these functions and initiate some of the cellular responses to altered renal physiology. The current study uses two-dimensional liquid chromatography/mass spectrometry to compare brush border membrane vesicles isolated from rat renal cortex (BBMV(CTX)) and from purified proximal convoluted tubules (BBMV(PCT)). Both proteomic data and Western blot analysis indicate that the BBMV(CTX) contain apical membrane proteins from cortical cells other than the proximal tubule. This heterogeneity was greatly reduced in the BBMV(PCT). Proteomic analysis identified 193 proteins common to both samples, 21 proteins unique to BBMV(CTX), and 57 proteins unique to BBMV(PCT). Spectral counts were used to quantify relative differences in protein abundance. This analysis identified 42 and 50 proteins that are significantly enriched (p values <or=0.001) in the BBMV(CTX) and BBMV(PCT), respectively. These data were validated by measurement of gamma-glutamyltranspeptidase activity and by Western blot analysis. The combined results establish that BBMV(PCT) are primarily derived from the proximal convoluted tubule (S1 and S2 segments), whereas BBMV(CTX) include proteins from the proximal straight tubule (S3 segment). Analysis of functional annotations indicated that BBMV(PCT) are enriched in mitochondrial proteins and enzymes involved in glucose and organic acid metabolism. Thus the current study reports a detailed proteomic analysis of the brush-border membrane of the rat renal proximal convoluted tubule and provides a database for future hypothesis-driven research.
Scott J Walmsley; Corey Broeckling; Ann Hess; Jessica Prenni; Norman P Curthoys
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, N.I.H., Extramural     Date:  2010-03-10
Journal Detail:
Title:  American journal of physiology. Renal physiology     Volume:  298     ISSN:  1522-1466     ISO Abbreviation:  Am. J. Physiol. Renal Physiol.     Publication Date:  2010 Jun 
Date Detail:
Created Date:  2010-05-21     Completed Date:  2010-06-14     Revised Date:  2013-05-30    
Medline Journal Info:
Nlm Unique ID:  100901990     Medline TA:  Am J Physiol Renal Physiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  F1323-31     Citation Subset:  IM    
Cell and Molecular Biology Program, Colorado State University, Fort Collins, Colorado 80523-1870, USA.
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MeSH Terms
Biological Markers / analysis
Blotting, Western
Centrifugation, Density Gradient
Chromatography, Liquid
Databases, Protein
Kidney Cortex / chemistry*
Kidney Tubules, Proximal / chemistry*
Membrane Proteins / isolation & purification*
Microvilli / chemistry
Proteomics* / methods
Rats, Sprague-Dawley
Reproducibility of Results
Tandem Mass Spectrometry
gamma-Glutamyltransferase / isolation & purification
Grant Support
Reg. No./Substance:
0/Biological Markers; 0/Membrane Proteins; EC

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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