Document Detail


Proteomic analysis of V-ATPase-rich cells harvested from the kidney and epididymis by fluorescence-activated cell sorting.
MedLine Citation:
PMID:  20181927     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Proton-transporting cells are located in several tissues where they acidify the extracellular environment. These cells express the vacuolar H(+)-ATPase (V-ATPase) B1 subunit (ATP6V1B1) in their plasma membrane. We provide here a comprehensive catalog of the proteins that are expressed in these cells, after their isolation by enzymatic digestion and fluorescence-activated cell sorting (FACS) from transgenic B1-enhanced green fluorescent protein (EGFP) mice. In these mice, type A and B intercalated cells and connecting segment cells of the kidney, and narrow and clear cells of the epididymis, which all express ATP6V1B1, also express EGFP, while all other cell types are negative. The proteome of renal and epididymal EGFP-positive (EGFP(+)) cells was identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and compared with their respective EGFP-negative (EGFP(-)) cell populations. A total of 2,297 and 1,564 proteins were detected in EGFP(+) cells from the kidney and epididymis, respectively. Out of these proteins, 202 and 178 were enriched by a factor greater than 1.5 in EGFP(+) cells compared with EGFP(-) cells, in the kidney and epididymis respectively, and included subunits of the V-ATPase (B1, a4, and A). In addition, several proteins involved in intracellular trafficking, signaling, and cytoskeletal dynamics were identified. A novel common protein that was enriched in renal and epididymal EGFP(+) cells is the progesterone receptor, which might be a potential candidate for the regulation of V-ATPase-dependent proton transport. These proteomic databases provide a framework for comprehensive future analysis of the common and distinct functions of V-ATPase-B1-expressing cells in the kidney and epididymis.
Authors:
Nicolas Da Silva; Trairak Pisitkun; Clémence Belleannée; Lance R Miller; Raoul Nelson; Mark A Knepper; Dennis Brown; Sylvie Breton
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, N.I.H., Intramural     Date:  2010-02-24
Journal Detail:
Title:  American journal of physiology. Cell physiology     Volume:  298     ISSN:  1522-1563     ISO Abbreviation:  Am. J. Physiol., Cell Physiol.     Publication Date:  2010 Jun 
Date Detail:
Created Date:  2010-05-21     Completed Date:  2010-06-14     Revised Date:  2011-07-28    
Medline Journal Info:
Nlm Unique ID:  100901225     Medline TA:  Am J Physiol Cell Physiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  C1326-42     Citation Subset:  IM    
Affiliation:
Harvard Medical School, Boston, MA, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Separation*
Chromatography, Liquid
Databases, Protein
Epididymis / cytology,  enzymology*
Flow Cytometry*
Gene Expression Regulation
Genotype
Green Fluorescent Proteins / biosynthesis,  genetics
Kidney / cytology,  enzymology*
Male
Mice
Mice, Transgenic
Phenotype
Promoter Regions, Genetic
Proteomics* / methods
RNA, Messenger / metabolism
Receptors, Progesterone / genetics,  metabolism
Tandem Mass Spectrometry
Vacuolar Proton-Translocating ATPases / genetics,  metabolism*
Grant Support
ID/Acronym/Agency:
DK-38452/DK/NIDDK NIH HHS; DK-42956/DK/NIDDK NIH HHS; DK-43341/DK/NIDDK NIH HHS; DK-57521/DK/NIDDK NIH HHS; HD-40793/HD/NICHD NIH HHS
Chemical
Reg. No./Substance:
0/RNA, Messenger; 0/Receptors, Progesterone; 0/enhanced green fluorescent protein; 147336-22-9/Green Fluorescent Proteins; EC 3.6.1.-/Atp6v1b1 protein, mouse; EC 3.6.1.-/Vacuolar Proton-Translocating ATPases
Comments/Corrections
Comment In:
Am J Physiol Cell Physiol. 2010 Jun;298(6):C1303-4   [PMID:  20237146 ]

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


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