Binding Interaction Between Prazosin and Immobilized Receptor by Frontal Analysis

Abstract

Binding interaction between ligand and G-protein coupled receptor plays an important role in the several steps of exertion of therapeutic effect by a drug and has become increasingly interesting to pharmacists, chemists, biologists and companies. a 1 -Adrenoceptor (a 1 -AR) was purified from cell line which stably expressed a 1 -AR and evenly immobilized on the surface of macroporous silica gel to prepare a novel stationary phase for investigating the interaction between drug and receptor. Control drugs of a 1 -AR including phentolamine, terazosin and urapidil were used to characterize the retention properties of the stationary phase containing the receptor. Further work was performed to calculate the binding sites and association constant of prazosin binding to the immobilized receptor. The results presented a value of 4.55 9 10 -7 M -1 for binding sites and of 2.2 9 10 4 M for the association constant during the interaction between prazosin and a 1 -AR. The proposed affinity method was stable at least in seven consecutive days, as well as had the primary bioactivities of recognizing and binding the ligand, providing an alternative for representing the interaction between drug and functional protein.

Identification of bioactive compounds in Shaoyao‐Gancao decoction using β2‐adrenoceptor affinity chromatography

“…Subsequently, the purified β 2 ‐AR protein was immobilized on the surface of a silica gel, according to a previous report . Briefly, the aminopropyl gel was activated by N,N ′‐carbonyldiimidazole in acetonitrile and loaded with the purified β 2 ‐AR.…”

“…The affinity constants and the numbers of binding sites of separated compounds that bound to the β 2 ‐AR were examined by frontal analysis, as previously reported . This assay is based on the shifting of the mean position of the breakthrough curves to shorter breakthrough times while infusing the immobilized β 2 ‐AR column with increased concentrations of compounds.…”

“…The affinity constants and the numbers of binding sites of separated compounds that bound to the β 2 -AR were examined by frontal analysis, as previously reported [21,22]. This assay is Journal of Separation Science…”

Development and evaluation of poly adenosine 5′‐diphosphate‐ribose polymerase 1 immobilization‐based receptor chromatography

“…In previous work, Zhao et al. immobilized β 2 ‐adrenoceptor ( β 2 ‐AR), an important drug target for respiratory diseases, onto macroporous silica gel by various immobilization methods [15–18]. The affinity columns were used to screen the bioactive compounds from TCMs, such as Siraitia grosvenorii [19] and Rhodiola crenulate [20].…”

Screening Bioactive Compounds of Siraitia grosvenorii by Immobilized β2-Adrenergic Receptor Chromatography and Druggability Evaluation

“…Although their application is successful in many cases, such methods need further improvement because most of the current methods concentrate on the transporting protein like serum albumin but much less on the largest drug targets such as G-protein-coupled receptors. Taking inspiration from the pioneer work by Wainer et al (Wainer et al, 1999; Moaddel and Wainer, 2006; Krzysztof et al, 2010), we have developed a series of immobilized G-protein-coupled receptor-based chromatographic methods (Zhao et al, 2010; Zhao et al, 2012; Zeng et al, 2018). Broad application and validation of these methods in real sample are required to make them a powerful tool for screening bioactive compounds of TCM.…”