AJP - Renal AJP: Cell Physiology
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Am J Physiol Renal Physiol 296: F204-F211, 2009. First published October 8, 2008; doi:10.1152/ajprenal.90473.2008
0363-6127/09 $8.00
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
296/1/F204    most recent
90473.2008v1
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 Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Schoeber, J. P. H.
Right arrow Articles by Bindels, R. J. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Schoeber, J. P. H.
Right arrow Articles by Bindels, R. J. M.

INNOVATIVE METHODOLOGY

Conditional fast expression and function of multimeric TRPV5 channels using Shield-1

Joost P. H. Schoeber,1,* Stan F. J. van de Graaf,2,* Kyu Pil Lee,1 Hanneke G. M. Wittgen,1 Joost G. J. Hoenderop,1 and René J. M. Bindels1

1Department of Physiology, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, Nijmegen; and 2Department of Metabolic and Endocrine Diseases and Netherlands Metabolomics Centre, UMC Utrecht, Utrecht, The Netherlands

Submitted 7 August 2008 ; accepted in final form 6 October 2008

A recently described novel controllable method to regulate protein expression is based on a mutated FK506-binding protein-12 (mtFKBP) that is unstable and rapidly degraded in mammalian cells. This instability can be conferred to other proteins directly fused to mtFKBP. Binding of a synthetic cell-permeant ligand (Shield-1) to mtFKBP reverses the instability, allowing conditional expression of mtFKBP-fused proteins. We adapted this strategy to study multimeric plasma membrane proteins using the ion channel TRPV5 as model protein. mtFKBP-TRPV5 forms functional ion channels and its expression can be controlled in a time- and dose-dependent fashion using Shield-1. Moreover, in the presence of Shield-1, mtFKBP-TRPV5 formed heteromultimeric channels with untagged TRPV5, which were codegraded upon washout of Shield-1, providing a strategy to study multimeric plasma membrane protein complexes without the need to destabilize all individual subunits.

conditional protein expression; inducible system; ion channels; heteromultimerization


Address for reprint requests and other correspondence: R. J. Bindels, Dept. of Physiology (286), Nijmegen Centre for Molecular Life Sciences, Radboud Univ. Nijmegen Medical Centre, Nijmegen 6500 HB, The Netherlands (e-mail: R.Bindels{at}ncmls.ru.nl)






HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Visit Other APS Journals Online
Copyright © 2009 by the American Physiological Society.