A fast affinity extraction methodology for rapid screening of bioactive compounds specifically binding to beta2-adrenergic receptor from Xie-Bai-San

摘要

Despite promising progress in series of techniques for drug discovery, the number of approved drugs has decreased in recent decades due to the lack of high efficient methods for candidate screening. In this work, immobilized beta2-adrenergic receptor was utilized to develop a new method for screening bioactive compounds from complex prescription, involving oriented immobilization of beta2-adrenergic receptor through diazonium reaction, the fast affinity extraction of bioactive compounds from Xie-Bai-San and the identification of these compounds by time of flight mass spectrometry. Valine, 4-hydroxyl resveratrol and kukoamine B were identified as the bioactive compounds specifically binding to beta2-adrenergic receptor in Xie-Bai-San. The binding of these compounds to the receptor occurred on four amino acid residues, viz., Asp 113, Ser 204, Ser 207, and Phe 290 in the crystal structure of beta2-adrenergic receptor. Hydrogen bond and hydrophobic force were believed to drive the binding of the compounds to beta2-adrenergic receptor on these residues. The results indicated that the proposed methodology has advantages on rapid, high throughput, and specific analysis, meanwhile, it has the capacity to probe the binding sites of the screened bioactive compounds. Fast affinity extraction methodology is expected to become a powerful alternative, which is superior in rapid screening of bioactive compounds from complex matrices such as traditional Chinese medicine.