Document Detail


Characterization of an extended receptive ligand repertoire of the human olfactory receptor OR17-40 comprising structurally related compounds.
MedLine Citation:
PMID:  16539658     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Molecular properties of odorant compounds essential for activation of the human olfactory receptor hOR17-40 were investigated using a collection of 23 variants of its cognate ligand helional. Coupling receptor activation to an optically detectable intracellular Ca(2+) ion flux allowed dose-dependent screening of different odorant molecules in human embryonic kidney (HEK)293 cells. We found an extended collection of activating ligands and provide first evidence for hOR17-40-specific antagonists. The C-terminal fusion of enhanced green fluorescent protein to the hOR17-40 retained full receptor function and permitted the selection of cells with defined receptor expression levels, which was an essential step for optimizing our screening protocol. Interestingly, cells with a low EGFP fluorescence intensity exhibited efficient hOR17-40 cell surface targeting and odorant-evoked signal transduction; in contrast, highly fluorescent cells displayed mainly incorrectly targeted, intracellular receptors. Fluorescence-activated cell sorting was used to separate hOR17-40-expressing cells on the basis of their endogenous EGFP fluorescence intensity, thereby increasing the fraction of odorant-responsive cells to up to 80% of the total cell number.
Authors:
Valérie Jacquier; Horst Pick; Horst Vogel
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't     Date:  2006-03-15
Journal Detail:
Title:  Journal of neurochemistry     Volume:  97     ISSN:  0022-3042     ISO Abbreviation:  J. Neurochem.     Publication Date:  2006 Apr 
Date Detail:
Created Date:  2006-04-25     Completed Date:  2006-06-08     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985190R     Medline TA:  J Neurochem     Country:  England    
Other Details:
Languages:  eng     Pagination:  537-44     Citation Subset:  IM    
Affiliation:
Institute of Chemical Sciences and Engineering, Ecole Polytechnique Fédérale de Lausanne (EPFL), Switzerland.
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MeSH Terms
Descriptor/Qualifier:
Calcium / metabolism
Cell Line
Cloning, Molecular / methods
Dose-Response Relationship, Drug
Flow Cytometry / methods
Gene Expression / drug effects,  physiology*
Green Fluorescent Proteins / metabolism
Humans
Ligands
Models, Molecular
Odors
Protein Structure, Secondary / physiology
Receptors, Odorant / chemistry*,  metabolism*
Signal Transduction / drug effects
Stimulation, Chemical
Transfection / methods
Chemical
Reg. No./Substance:
0/Ligands; 0/Receptors, Odorant; 0/enhanced green fluorescent protein; 147336-22-9/Green Fluorescent Proteins; 7440-70-2/Calcium

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


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