Biochemical and Biophysical Research Communications, Vol.494, No.3-4, 634-640, 2017
Investigation on temperature-induce conformational change of immobilized beta(2) adrenergic receptor
The beta(2) adrenergic receptor (beta(2)-AR) is a prototypical family A G protein-coupled receptor (GPCR) and an excellent model system for studying the mechanism of GPCR activation. Purified beta(2)-AR was immobilized on macroporous silica gel to obtain liquid chromatographic stationary phase. The resulting phase was packed into a stainless steel column (4.6 x 50 mm, 7 mu m) and used for on-line chromatographic system. When column oven temperature increased from 20.0 degrees C to 40.0 degrees C, uncomplete separate chromatographic peaks of ephedrine and pseudoephedrine as receptor conformational probe were gradually merged into one peak, meanwhile retention time and resolution of the probes were reduced correspondingly, which suggested that temperature could regulate protein conformation. Temperature induced conformational change of immobilized beta(2)-AR, especially changes at higher temperatures, indicated that constructed receptor chromatography could simulate fever disease state of human body and clarify receptor conformation change at pathological condition. At the same time this study could also provide new ideas for screening active components in pathological conditions. (C) 2017 Elsevier Inc. All rights reserved.
Keywords:beta(2) adrenergic receptor;Temperature-induced;Protein conformation;Receptor chromatography;Ephedrine and pseudoephedrine