title: Confirming therapeutic target of protopine using immobilized β2-adrenoceptor coupled with site-directed molecular docking and the target-drug interaction by frontal analysis and injection amount-dependent method.
abstract: Drug-protein interaction analysis is pregnant in designing new leads during drug discovery. We prepared the stationary phase containing immobilized β2-adrenoceptor (β2-AR) by linkage of the receptor on macroporous silica gel surface through N,N'-carbonyldiimidazole method. The stationary phase was applied in identifying antiasthmatic target of protopine guided by the prediction of site-directed molecular docking. Subsequent application of immobilized β2-AR in exploring the binding of protopine to the receptor was realized by frontal analysis and injection amount-dependent method. The association constants of protopine to β2-AR by the 2 methods were (1.00 ± 0.06) × 105M-1and (1.52 ± 0.14) × 104M-1. The numbers of binding sites were (1.23 ± 0.07) × 10-7M and (9.09 ± 0.06) × 10-7M, respectively. These results indicated that β2-AR is the specific target for therapeutic action of protopine in vivo. The target-drug binding occurred on Ser169in crystal structure of the receptor. Compared with frontal analysis, injection amount-dependent method is advantageous to drug saving, improvement of sampling efficiency, and performing speed. It has grave potential in high-throughput drug-receptor interaction analysis.
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an, China.
authors.forename: Guangxin
authors.initials: G
authors.lastname: Liu
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an, China.
authors.forename: Pei
authors.initials: P
authors.lastname: Wang
authors.affiliation: Drug Certificate Center, Shaanxi Food and Drug Administration, Xi'an, China.
authors.forename: Chan
authors.initials: C
authors.lastname: Li
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an, China.
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Sun
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an, China.
authors.forename: Xinfeng
authors.identifier: 0000-0001-8824-738X
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an, China.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Zheng
mesh_terms: D053119:Benzophenanthridines / Q000737:chemistry / Q000494:pharmacology*; D001600:Berberine Alkaloids / Q000737:chemistry / Q000494:pharmacology*; D001665:Binding Sites / Q000187:drug effects; D002846:Chromatography, Affinity; D004347:Drug Interactions; D006801:Humans; D007093:Imidazoles / Q000737:chemistry; D062105:Molecular Docking Simulation; D058990:Molecular Targeted Therapy; D011485:Protein Binding / Q000187:drug effects; D018343:Receptors, Adrenergic, beta-2 / Q000737:chemistry* / Q000187:drug effects
chemical_list: D053119:Benzophenanthridines; D001600:Berberine Alkaloids; D007093:Imidazoles; D018343:Receptors, Adrenergic, beta-2; C006900:N,N-carbonyldiimidazole; C009093:protopine
keywords: drug-receptor interaction; frontal analysis; immobilized β2-adrenoceptor; protopine