Assessment of a fully active class A G protein-coupled receptor isolated from in vitro folding.

摘要: We provide a protocol for the preparation of fully active Y2 G protein-coupled receptors (GPCRs). Although valuable target pharmaceutical research, information about structure and dynamics these molecules remains limited due to difficulty in obtaining sufficient amounts homogeneous vitro studies. Recombinant expression GPCRs as inclusion bodies provides highest protein yields at lowest costs. But this strategy can only successfully be applied if subsequent folding results high yield fraction isolated from nonactive form. Here, we followed that large quantities human neuropeptide Y receptor type 2 determined before after ligand affinity chromatography using radioligand binding assay. Directly folding, achieved proportion ∼25% receptor. This value could increased ∼96% ligand...