title: HDL-like discs for assaying membrane proteins in drug discovery.
abstract: To broaden the use of the recombinant high-density lipoprotein (rHDL) approach to the characterization of lead compounds, we investigated the pharmacology of the human beta-2-adrenoceptor in nanolipid bilayers (rHDL) with a broad set of beta-adrenoceptor antagonists. To that end, we developed a homogeneous copper-chelate scintillation proximity binding assay (SPA) in order to compare receptor-ligand binding affinities before and after reconstitution into rHDLs. Our results clearly show that the beta-2-adrenoceptor reconstituted in rHDLs display the same pharmacology as that in cell membranes and that rHDLs can be used not only to measure affinities for a range of ligands but also to study binding kinetics.
authors.affiliation: Novartis Institutes for BioMedical Research, Novartis Campus, 4056 Basel, Switzerland.
authors.forename: Cédric
authors.initials: C
authors.lastname: Fiez-Vandal
authors.forename: Lukas
authors.initials: L
authors.lastname: Leder
authors.forename: Felix
authors.initials: F
authors.lastname: Freuler
authors.forename: David
authors.initials: D
authors.lastname: Sykes
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Charlton
authors.forename: Sandra
authors.initials: S
authors.lastname: Siehler
authors.forename: Ulrich
authors.initials: U
authors.lastname: Schopfer
authors.forename: Myriam
authors.initials: M
authors.lastname: Duckely
mesh_terms: D058672:Adrenergic beta-2 Receptor Antagonists / Q000378:metabolism; D000818:Animals; D016632:Apolipoprotein A-I / Q000378:metabolism; D055808:Drug Discovery; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D008075:Lipoproteins, HDL / Q000378:metabolism*; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D018343:Receptors, Adrenergic, beta-2 / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D058672:Adrenergic beta-2 Receptor Antagonists; D016632:Apolipoprotein A-I; D008051:Lipid Bilayers; D008075:Lipoproteins, HDL; D018343:Receptors, Adrenergic, beta-2; D011994:Recombinant Proteins