title: Characterization of radioligand binding to a transmembrane receptor reconstituted into Lipobeads.
abstract: Lipobeads are hydrogel beads surrounded by a lipid bilayer membrane and have been developed to act as a cell analogue. The FLAG-tagged M(2) muscarinic receptor was incorporated onto the surface of the Lipobead by incubating pre-Lipobeads with proteoliposomes containing the receptor. Receptors reconstituted onto the surface of the Lipobeads were functional in that they bound the antagonists quinuclidinylbenzilate and scopolamine with characteristic muscarinic affinities. This demonstrates the feasibility of using Lipobeads to study the binding properties of the M(2) muscarinic receptor and offers a promising approach to the study of transmembrane protein biology in general.
authors.affiliation: Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, Ont., Canada M5S 2S2.
authors.forename: Paul S-H
authors.initials: PS
authors.lastname: Park
authors.forename: Charlene C
authors.initials: CC
authors.lastname: Ng
authors.forename: Sean
authors.initials: S
authors.lastname: Buck
authors.forename: James W
authors.initials: JW
authors.lastname: Wells
authors.forename: Yu-Ling
authors.initials: YL
authors.lastname: Cheng
authors.forename: Peter S
authors.initials: PS
authors.lastname: Pennefather
mesh_terms: D000345:Affinity Labels; D000818:Animals; D002460:Cell Line; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D008863:Microspheres; D018727:Muscarinic Antagonists / Q000378:metabolism / Q000494:pharmacology; D011510:Proteolipids / Q000378:metabolism; D011813:Quinuclidinyl Benzilate / Q000378:metabolism / Q000494:pharmacology; D011869:Radioligand Assay; D043585:Receptor, Muscarinic M2 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012601:Scopolamine / Q000378:metabolism / Q000494:pharmacology; D018411:Spodoptera / Q000166:cytology
chemical_list: D000345:Affinity Labels; D008051:Lipid Bilayers; D018727:Muscarinic Antagonists; D011510:Proteolipids; D043585:Receptor, Muscarinic M2; C020485:proteoliposomes; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D011813:Quinuclidinyl Benzilate; D012601:Scopolamine