| Studying on the interaction between small moleculars and receptors plays a very important role in the mechanism of drugs,new drug development,drug screening and so on.β2-adrenoceptor belongs to G-protein coupled receptors,who mainly distributed in smooth muscles and it is mainly used for the treatment of respiratory diseases. In the isolated environment,we need pureβ2-AR to study its structure and the interaction between small moleculars andβ2-AR,but the technology of purification forβ2-AR has not been solved so far.So the key link to purificateβ2-AR is its falling from the organization of the cells to the solution.This paper mainly focus onβ2-adrenoceptor,so the main results are as follows:Firstly, after comparing five different kinds of detergents, including Deoxycholic Acid Sodium Salt,SDS Sodium dodecyl sulfate(SDS),Sodium taurocholate,Cholic Acid Soldium Salt and the admixture which consist of digitonin/sodium cocholate/CHAPS.Based on SDS-PAGE,we found the most effectivel solubilized of purificateβ2-adrenocepters from dog's lung is an admixture which consist of digitonin/sodium cocholate/CHAPS(mass ratio:1:5:2).Secondly, The binding sites and association constant of immobilized were determinated by the methods of frontal analysis and zonal elution.The results are as follows:Propranolol hydrochloride andβ2-adrenocepters has two types of binding sites, the total binding sites was 3.27×10-6M, association constants were 2.12×104M-1,0.01×104M-1;Clenbuterol hydrochloride andβ2-adrenocepters has only one type binding site, the binding sites of clenbuterol hydrochloride withβ2-adrenocepters on the biochromatographic column was 1.07×10-6M, the association constant was 0.53×104M-1.the energy distribution of the latter one is homogeneous,but the former one is opposite.
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