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Functional reconstitution of purified metabotropic glutamate receptor expressed in the fly eye
Author(s) -
Eroglu Çagla,
Cronet Philippe,
Panneels Valérie,
Beaufils Philippe,
Sinning Irmgard
Publication year - 2002
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.1093/embo-reports/kvf088
Subject(s) - g protein coupled receptor , metabotropic glutamate receptor , metabotropic glutamate receptor 6 , biology , metabotropic receptor , microbiology and biotechnology , receptor , biochemistry , metabotropic glutamate receptor 5 , drosophila melanogaster , metabotropic glutamate receptor 1 , metabotropic glutamate receptor 7 , metabotropic glutamate receptor 4 , glutamate receptor , gene
G‐protein‐coupled receptors (GPCRs) form one of the largest superfamilies of membrane proteins. Obtaining high yields of GPCRs remains one of the major factors limiting a detailed understanding of their structure and function. Photoreceptor cells (PRCs) contain extensive stacks of specialized membranes where high levels of rhodopsins are naturally present, which makes them ideal for the overexpression of GPCRs. We have generated transgenic flies expressing a number of GPCRs in the PRCs. Drosophila melanogaster metabotropic glutamate receptor (DmGluRA) expressed by this novel strategy was purified to homogeneity, giving at least 3‐fold higher yields than conventional baculovirus expression systems due to the higher membrane content of the PRCs. Pure DmGluRA was then reconstituted into liposomes of varying composition. Interestingly, glutamate binding was strictly dependent on the presence of ergosterol.