Summary of experimental strategy.

Abstract

<p>A, Testing stabilizing effects of detergents, ligands and lipids. •Efficient solubilization of the fusion CB₂ protein from <i>E. coli</i> membranes. Over 40 different detergents and mixtures of detergents were compared for their efficiency in solubilizing fusion CB₂ from membranes, and the detergent mixture that performed best was selected for a routine receptor purification protocol. •Optimization of the liposome-reconstitution procedure. A screening of lipid-solubilizing detergents was performed with a goal of maximizing the yield of functionally reconstituted receptor. •Application of the G protein activation test to the analysis of the structural stability of CB₂ in micelles following its reconstitution into liposomes. •Screening for stabilizers for CB₂ and characterization of stability of the purified receptor. (i)Stabilization of CB₂ in micelles by CHS, ligands and phospholipids. (ii)Ligand binding studies by solid-state NMR. (iii)Characterization of stability of CB₂ in lipid bilayers. B, Comparison of stabilizing effects of two lipids at two different temperatures.</p