Agonist-independent localization of the NOP receptor in detergent-resistant membrane rafts.
A lipid rafts/detergent-resistant membrane (DRM) fraction was prepared from recombinant HEK293 cells stably expressing the human NOP receptor fused to the green fluorescent protein EGFP (hNOPr-EGFP), and probed for the presence and functionality of the fusion protein. Fluorescence detection as well as immunoblotting with an anti-GFP antibody revealed that most of the fusion protein was recovered in the DRM fraction, wherein it mediated efficiently NOP-induced stimulation of GTPgamma(35)S binding. Recovery of hNOPr-EGFP in the DRM fraction was not affected had the cells been acutely or chronically exposed to NOP prior to detergent treatment. Therefore, in HEK cells, the NOP receptor localizes constitutively to DRMs wherein it retains ability to couple with hetero-trimeric G protein.
Butour
Jean-Luc
JL
Institut de Pharmacologie et de Biologie Structurale, UMR 5089, Centre National de la Recherche Scientifique, 205 route de Narbonne, 31077 Toulouse cédex 04, France.
Corbani
Maïthé
M
Meunier
Jean-Claude
JC
D002460:Cell Line / Q000187:drug effects; D003902:Detergents / Q000494:pharmacology; D004351:Drug Resistance / Q000502:physiology; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006801:Humans; D007668:Kidney / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D021962:Membrane Microdomains / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D017830:Octoxynol / Q000494:pharmacology*; D011957:Receptors, Opioid / Q000187:drug effects / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D000094942:Nociceptin Receptor
D003902:Detergents; D011957:Receptors, Opioid; D011994:Recombinant Proteins; D017830:Octoxynol; D019204:GTP-Binding Proteins; D000094942:Nociceptin Receptor; C000722283:OPRL1 protein, human