Mutiresidue Detection of β Aeceptor Agonists in β2Adrenergic Receptor by Artificial Expression
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Abstract:
This study was directed to improve the activity of β2 adrenergic receptor and to identify the activity ofβ2 adrenergic receptor by artificial expression.We used molecular docking technology to separately model the interaction between β2adrenergic receptor and fifteen kinds ofβ receptor agonists. Based on the molecular structure of β2 adrenergic receptor, the β receptor agonists with best binding in molecular docking was selected, and the β2adrenergic receptor gene was synthesized and optimized. The recombinant expression plasmid was constructed to be transferred to Escherichia coil BL21(DE3), which expressed the active receptor protein successfully. SDS-PAGE of recombinant expression plasmid showed that theβ2 adrenergicreceptor protein was approximately 47 ku. The results of the receptor activity showed that the receptor protein was capable of specific reaction with Clenbuterol, ractopamine, and fenoterol respectively. OD values were 0.45, 0.32 and 0.36 respectively, and the binding ability of the receptor protein and β receptor agonists was consistent with the results of the activity identification basically. Testing curve shows that β2 adrenergic receptor protein have the ability to detect β receptor agonists. Using molecular docking technology, we successfully obtained the activeβ2adrenergic receptor protein, which laid the foundation for the application of β2adrenergic receptor to the multresidue detection of β receptor agonists.
