MCP Agilent Technologies
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/mcp.M600177-MCP200 on August 9, 2006.
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental Data
Right arrow All Versions of this Article:
M600177-MCP200v1
5/11/2131    most recent
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
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 Glossary
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 Yu, M.-J.
Right arrow Articles by Knepper, M. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Yu, M.-J.
Right arrow Articles by Knepper, M. A.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Molecular & Cellular Proteomics 5:2131-2145, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

LC-MS/MS Analysis of Apical and Basolateral Plasma Membranes of Rat Renal Collecting Duct Cells*,S

Ming-Jiun Yu{ddagger}, Trairak Pisitkun{ddagger}, Guanghui Wang§, Rong-Fong Shen§ and Mark A. Knepper{ddagger},¶

From the {ddagger} Laboratory of Kidney and Electrolyte Metabolism and § Proteomics Core Facility, NHLBI, National Institutes of Health, Bethesda, Maryland 20892

We used biotinylation and streptavidin affinity chromatography to label and enrich proteins from apical and basolateral membranes of rat kidney inner medullary collecting ducts (IMCDs) prior to LC-MS/MS protein identification. To enrich apical membrane proteins and bound peripheral membrane proteins, IMCDs were perfusion-labeled with primary amine-reactive biotinylation reagents at 2 °C using a double barreled pipette. The perfusion-biotinylated proteins and proteins bound to them were isolated with CaptAvidin-agarose beads, separated with SDS-PAGE, and sliced into continuous gel pieces for LC-MS/MS protein identification (LTQ, Thermo Electron Corp.). 17 integral and glycosylphosphatidylinositol (GPI)-linked membrane proteins and 44 non-integral membrane proteins were identified. Immunofluorescence confocal microscopy confirmed ACVRL1, H+/K+-ATPase {alpha}1, NHE2, and TauT expression in the IMCDs. Basement membrane and basolateral membrane proteins were biotinylated via incubation of IMCD suspensions with biotinylation reagents on ice. 23 integral and GPI-linked membrane proteins and 134 non-integral membrane proteins were identified. Analyses of non-integral membrane proteins preferentially identified in the perfusion-biotinylated and not in the incubation-biotinylated IMCDs revealed protein kinases, scaffold proteins, SNARE proteins, motor proteins, small GTP-binding proteins, and related proteins that may be involved in vasopressin-stimulated AQP2, UT-A1, and ENaC regulation. A World Wide Web-accessible database was constructed of 222 membrane proteins (integral and GPI-linked) from this study and prior studies.

To whom correspondence should be addressed: National Institutes of Health, Bldg. 10, Rm. 6N260, 10 Center Dr., MSC-1603, Bethesda, MD 20892-1603. Tel.: 301-496-3064; Fax: 301-402-1443; E-mail: knep{at}helix.nih.gov


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 Am. J. Physiol. Renal Physiol.Home page
C.-L. Chou, M.-J. Yu, E. M. Kassai, R. G. Morris, J. D. Hoffert, S. M. Wall, and M. A. Knepper
Roles of basolateral solute uptake via NKCC1 and of myosin II in vasopressin-induced cell swelling in inner medullary collecting duct
Am J Physiol Renal Physiol, July 1, 2008; 295(1): F192 - F201.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Renal Physiol.Home page
Y.-J. Lee, H.-J. Choi, J.-S. Lim, J.-H. Earm, B.-H. Lee, I.-S. Kim, J. Frokiaer, S. Nielsen, and T.-H. Kwon
A novel method of ligand peptidomics to identify peptide ligands binding to AQP2-expressing plasma membranes and intracellular vesicles of rat kidney
Am J Physiol Renal Physiol, July 1, 2008; 295(1): F300 - F309.
[Abstract] [Full Text] [PDF]

Home page
 Physiol. GenomicsHome page
P. Uawithya, T. Pisitkun, B. E. Ruttenberg, and M. A. Knepper
Transcriptional profiling of native inner medullary collecting duct cells from rat kidney
Physiol Genomics, January 17, 2008; 32(2): 229 - 253.
[Abstract] [Full Text] [PDF]

Home page
 PhysiologyHome page
T. Pisitkun, J. D. Hoffert, M.-J. Yu, and M. A. Knepper
Tandem Mass Spectrometry in Physiology
Physiology, December 1, 2007; 22(6): 390 - 400.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Journal of Biological Chemistry 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2006 by the American Society for Biochemistry and Molecular Biology.