Solubilization and functional reconstitution of human neuropeptide FF2 receptors.
Neuropeptide FF (NPFF, FLFQPQRFamide) receptors modulate endogenous opioid functions. Here, we report the solubilization of the human NPFF(2) receptor expressed in Chinese hamster ovary (CHO) cells by the zwitterionic detergent Chaps. Chaps solubilization resulted in the abolishment of specific agonist binding activity, which was restored by a polyethylene glycol (PEG) precipitation method. Reincorporation after the precipitation step into liposomes made of endogenous lipids issued from CHO membranes or exogenous lipids significantly enhanced the specific agonist binding activity and G-protein coupling. This method of solubilization and lipid reconstitution could be useful for studies of NPFF receptors.
Talmont
Frank
F
Institut de Pharmacologie et de Biologie Structurale, CNRS/Université de Toulouse, UMR 5089, 205 route de Narbonne, Toulouse Cedex, France.
Mollereau
Catherine
C
Muller
Isabelle
I
Zajac
Jean-Marie
JM
D000818:Animals; D016466:CHO Cells; D002793:Cholic Acids / Q000494:pharmacology; D006224:Cricetinae; D003412:Cricetulus; D003902:Detergents / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D018013:Receptors, Neuropeptide / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012995:Solubility
D002793:Cholic Acids; D003902:Detergents; D018013:Receptors, Neuropeptide; C083929:neuropeptide FF receptor; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate