title: Target-directed screening of the bioactive compounds specifically binding to β₂-adrenoceptor in Semen brassicae by high-performance affinity chromatography.
abstract: The bioactive ingredients in Semen sinapis were rapidly screened by immobilized β2-adrenoceptor (β2-AR) and target-directed molecular docking. The methods involved the attachment of β2-AR using any amino group in the receptor, the simultaneous separation and identification of the retention compounds by high-performance affinity chromatography; the binding mechanism of the interesting compound to the receptor was investigated by zonal elution and molecular docking. Sinapine in Semen sinapis was proved to be the bioactive compound that specifically binds to the immobilized receptor. The association constant of sinapine to β2-AR was determined to be 1.36 × 10(5)  M(-1) with a value of 1.27 × 10(-6)  M for the number of binding sites. Ionic bond was believed to be the driving force during the interaction between sinapine and β2-AR. It is possible to become a powerful alternative for rapid screening of bioactive compounds from a complex matrix such as traditional Chinese medicine and further investigation on the drug-receptor interaction.
authors.affiliation: College of Life Sciences, Northwest University, Xi'an, 710069, China.
authors.forename: Yuxin
authors.initials: Y
authors.lastname: An
authors.affiliation: Shaanxi Institute for Food and Drug Control, Xi'an, 710065, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Li
authors.affiliation: College of Life Sciences, Northwest University, Xi'an, 710069, China.
authors.forename: Huanmei
authors.initials: H
authors.lastname: Sun
authors.affiliation: The Affiliated High School of Northwest University, Xi'an, 710075, China.
authors.forename: Wenhai
authors.initials: W
authors.lastname: Bian
authors.affiliation: Institute of Vascular Medicine, Third Hospital and Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education, Peking University, Beijing, 100083, China.
authors.forename: Zijian
authors.initials: Z
authors.lastname: Li
authors.affiliation: Institute of Vascular Medicine, Third Hospital and Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education, Peking University, Beijing, 100083, China.
authors.forename: Youyi
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Life Sciences, Northwest University, Xi'an, 710069, China.
authors.forename: Xinfeng
authors.initials: X
authors.lastname: Zhao
authors.affiliation: College of Life Sciences, Northwest University, Xi'an, 710069, China.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Zheng
mesh_terms: D002794:Choline / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods; D008516:Medicine, Chinese Traditional; D062105:Molecular Docking Simulation / Q000379:methods; D064209:Phytochemicals / Q000737:chemistry / Q000378:metabolism*; D011943:Receptors, Adrenergic, beta / Q000378:metabolism*; D031226:Sinapis / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D064209:Phytochemicals; D011943:Receptors, Adrenergic, beta; C005685:sinapine; D002794:Choline
keywords: Semen brassicae; drug screening; high-performance affinity chromatography; immobilized β2-AR; molecular docking