Covalent immobilization of beta2 adrenergic receptor through trans-methylation reaction by SNAP-tag and its application in anti-asthmatic compound screening from Raphani Semen

J Pharm Biomed Anal. 2022 Sep 20;219:114952. doi: 10.1016/j.jpba.2022.114952. Epub 2022 Jul 19.


Beta2-adrenergic receptor (β2-AR) is believed as an attractive target for anti-asthmatic drugs. Its crystal structure and pharmacological activity have been clearly investigated. Yet the number of the approved anti-asthmatic drugs has declined in recent years. This work reports on the preparation of an immobilized β2-AR column through the specific trans-methylation reaction between SNAP tag and the benzyl-guanine derivative and application in anti-asthmatic compound screening from Raphani Semen. The characterization of the immobilized β2-AR was performed by scanning electron microscopy (SEM) and receptor-ligand interaction analysis by chromatographic methods. SEM analysis showed that the receptor has been successfully coated on the surface of PEGA amino microspheres. Binding constants of salbutamol and terbutaline calculated from frontal analysis within the temperature range of 10-30 ℃ confirmed the feasibility of the method in a thermodynamic viewpoint. Hydrogen bond was verified as the main driving force for drug-receptor interaction analysis. Sinapine was identified as the potential bioactive compound in Raphani Semen that specifically bind with β2-AR with a specific binding site of Ser 207. Taking together, the immobilized β2-AR column is promising in exploring drug-protein interaction analysis and anti-asthmatic drug screening.

Keywords: Affinity chromatography; Beta2 adrenergic receptor; Bioactive compounds screening; Raphani Semen; Sinapine; Transmethylation reaction.

MeSH terms

  • Anti-Asthmatic Agents*
  • Binding Sites
  • Methylation
  • Protein Binding
  • Receptors, Adrenergic, beta-2 / metabolism
  • Seeds / metabolism


  • Anti-Asthmatic Agents
  • Receptors, Adrenergic, beta-2