Thermostability of a recombinant G protein-coupled receptor expressed at high level in mammalian cell culture.

ABSTRACT: Rational design of pharmaceutical drugs targeting integral membrane G protein-coupled receptors (GPCR) requires thorough understanding of ligand binding and mechanism of activation through high resolution structural studies of purified proteins. Due to inherent conformational flexibility of GPCR, stabilization of these proteins solubilized from cell membranes into detergents is a challenging task. Here, we take advantage of naturally occurring post-translational modifications for stabilization of purified GPCR in detergent micelles. The recombinant cannabinoid CB₂ receptor was expressed at high yield in Expi293F mammalian cell cultures, solubilized and purified in Façade detergent. We report superior stability of the mammalian cell-expressed receptor compared to its E. coli-expr

SUBMITTER: Yeliseev A 

PROVIDER: S-EPMC7546613 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature