Document Detail

Subcellular imaging of dynamic protein interactions by bioluminescence resonance energy transfer.
MedLine Citation:
PMID:  17921204     Owner:  NLM     Status:  MEDLINE    
Despite the fact that numerous studies suggest the existence of receptor multiprotein complexes, visualization and monitoring of the dynamics of such protein assemblies remain a challenge. In this study, we established appropriate conditions to consider spatiotemporally resolved images of such protein assemblies using bioluminescence resonance energy transfer (BRET) in mammalian living cells. Using covalently linked Renilla luciferase and yellow fluorescent proteins, we depicted the time course of dynamic changes in the interaction between the V2-vasopressin receptor and beta-arrestin induced by a receptor agonist. The protein-protein interactions were resolved at the level of subcellular compartments (nucleus, plasma membrane, or endocytic vesicules) and in real time within tens-of-seconds to tens-of-minutes time frame. These studies provide a proof of principle as well as experimental parameters and controls required for high-resolution dynamic studies using BRET imaging in single cells.
Vincent Coulon; Martin Audet; Vincent Homburger; Joël Bockaert; Laurent Fagni; Michel Bouvier; Julie Perroy
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2007-10-05
Journal Detail:
Title:  Biophysical journal     Volume:  94     ISSN:  1542-0086     ISO Abbreviation:  Biophys. J.     Publication Date:  2008 Feb 
Date Detail:
Created Date:  2008-01-10     Completed Date:  2008-02-05     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  0370626     Medline TA:  Biophys J     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1001-9     Citation Subset:  IM    
Institut de Génomique fonctionnelle, CNRS UMR5203, INSERM U661, University of Montpellier, Montpellier, France.
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MeSH Terms
Cell Line
Fluorescence Resonance Energy Transfer / methods*
Kidney / metabolism*
Luminescent Measurements
Luminescent Proteins / metabolism*
Microscopy, Fluorescence / methods*
Protein Interaction Mapping / methods*
Subcellular Fractions / metabolism*,  ultrastructure*
Reg. No./Substance:
0/Luminescent Proteins

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

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