1 research outputs found
Reliable Analysis of the Interaction between Specific Ligands and Immobilized Beta-2-Adrenoceptor by Adsorption Energy Distribution
Although
a comparatively robust method, immobilized protein-based
techniques have displayed limited precision and inconsistent results
due to a lack of strategy for the accurate selection of drug adsorption
models on the protein surface. We generated the adsorption data of
three drugs on immobilized beta-2-adrenoceptor (β<sub>2</sub>-AR) by frontal affinity chromatography–mass spectrometry
(FAC-MS) and site-specific competitive FAC-MS. Using adsorption energy
distribution (AED) calculations, we achieved the best adsorption models
for the binding of salbutamol, terbutaline, and pseudoephedrine to
immobilized β<sub>2</sub>-AR. The Langmuir model proved to be
desirable for describing the adsorptions of salbutamol and terbutaline
on immobilized β<sub>2</sub>-AR, while the bi-Langmuir model
was favorable to characterize the adsorption of pseudoephedrine on
the receptor. Relying on the accurate determination of association
constants, we presented an efficient approach for β<sub>2</sub>-AR ligand screening based on the loss of breakthrough time of an
indicator drug caused by the inclusion of competitive drugs in the
mobile phase. We concluded that the current strategy enables the reliable
and accurate analysis of G protein-coupled receptor (GPCR)–drug
interaction. The percentage change in the breakthrough time for drugs
can provide useful information for estimating their binding affinity
to the receptor. This approach builds a powerful platform for high-throughput
ligand screening