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Expression and functional analysis of olfactory receptors
H Breer1, J Krieger, C Meinken
1Institute of Physiology, University of Stuttgart-Hohenheim, Germany. physiol1@uni-hohenheim.de
Annals of the New York Academy of Sciences
|February 4, 1999
Summary
Researchers expressed olfactory receptor proteins in insect cells, demonstrating their ability to detect specific odorants. This confirms the function of these receptors in scent perception and provides a method for studying their interactions with various odor molecules.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- The olfactory system uses a vast array of seven-transmembrane domain receptors to detect diverse odorants.
- Direct evidence linking specific odorant receptors to odorant binding and signal transduction is needed.
Purpose of the Study:
- To confirm that identified genes encode functional odorant receptors.
- To demonstrate that these receptors can bind odorants and initiate signaling cascades.
- To establish a system for studying olfactory receptor ligand specificity.
Main Methods:
- Heterologous expression of olfactory receptor proteins using the baculovirus/Sf9 cell system.
- Stimulation of receptor-expressing cells with various odorants and measurement of second messenger responses.
- Production of olfactory receptor proteins in a bacterial system, solubilization, and reconstitution in liposomes for ligand binding assays.
Main Results:
- High-level expression of functional olfactory receptor proteins was achieved in Sf9 cells.
- Receptor-expressing cells showed significant, dose-dependent responses to odorant mixtures.
- Specific receptor subtypes exhibited selective, albeit broad, responses to distinct odorants.
- Reconstituted receptor proteins in liposomes demonstrated specific binding to odor ligands.
Conclusions:
- The study provides unequivocal proof that the expressed proteins are functional odorant receptors.
- The baculovirus/Sf9 cell system and bacterial expression system are effective for studying olfactory receptor function and ligand interactions.
- Olfactory receptors display selective but broad ligand specificity, contributing to odor discrimination.