The preparation, solubilisation and binding characteristics of a beta 2-adrenoceptor isolated from transfected Chinese hamster cells

The Analyst
S A MeenaghR F Witkamp

Abstract

beta 2-Adrenoceptor agonists (beta-agonists) are well known for their growth promoting and repartitioning effects in many species. Although the use of these compounds to increase muscle mass in stockfarming is prohibited within the EU, under directive 96/22/EC, significant illegal use still occurs. With legal and illegal synthesis of new structurally related compounds, the detection of traditional beta-agonists and new derivatives becomes increasingly problematical. This method describes the isolation and solubilisation of a beta 2-adrenoceptor from a transfected Chinese hamster ovary cell line, using the detergent digitonin. The solubilised receptor retained its activity and was isolated from the cell membrane at a concentration of 550 +/- 100 fmol mg-1 of solubilised protein. Competition analysis using the tritiated antagonist dihydroalprenolol revealed receptor affinity for five structurally different beta-agonists, with IC50 values ranging from 2.1 +/- 0.76 x 10(-7) M for salmeterol to 1.1 +/- 0.62 x 10(-5) M for ractopamine. This study has demonstrated that transfected cell lines with a high expression of beta 2-adrenoceptors are a convenient source of active receptor material. Solubilised beta 2-adrenoceptors could form t...Continue Reading

Citations

Apr 4, 2009·Journal of Chromatography. a·T F H Bovee, M G Pikkemaat
Apr 20, 2016·Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment·Toine F H BoveeRon L A P Hoogenboom

❮ Previous
Next ❯

Related Concepts

Related Feeds

Adrenergic Receptors: Trafficking

Adrenergic receptor trafficking is an active physiological process where adrenergic receptors are relocated from one region of the cell to another or from one type of cell to another. Discover the latest research on adrenergic receptor trafficking here.