Construction and application of H1R ligand screening materials based on SMA stabilization and his-tag covalent immobilization of membrane proteins.

Autor: Shan Y; School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, PR China., Xia Z; School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, PR China., An M; School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, PR China., Yang W; School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, PR China., Wang S; School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, PR China., Yang F; School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, PR China., He L; School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, PR China., Wang C; School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, PR China. Electronic address: chengwang@mail.xjtu.edu.cn., He H; School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, PR China. Electronic address: hehuaizhen@xjtu.edu.cn.
Jazyk: angličtina
Zdroj: Journal of chromatography. A [J Chromatogr A] 2024 Aug 16; Vol. 1729, pp. 465057. Date of Electronic Publication: 2024 Jun 06.
DOI: 10.1016/j.chroma.2024.465057
Abstrakt: The histamine H1 receptor (H1R) plays a pivotal role in allergy initiation and undergoes the necessity of devising a high-throughput screening approach centered on H1R to screen novel ligands effectively. This study suggests a method employing styrene maleic acid (SMA) extraction and His-tag covalent bonding to immobilize H1R membrane proteins, minimizing the interference of nonspecific proteins interference while preserving native protein structure and maximizing target exposure. This approach was utilized to develop a novel material for high-throughput ligand screening and implemented in cell membrane chromatography (CMC). An H1R-His-SMALPs/CMC model was established and its chromatographic performance (selectivity, specificity and lifespan) validated, demonstrating a significant enhancement in lifespan compared to previous CMC models. Subsequently, this model facilitated high-throughput screening of H1R ligands in the compound library and preliminary activity verification of potential H1R antagonists. Identification of a novel H1R antagonist laid the foundation for further development in this area.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024. Published by Elsevier B.V.)
Databáze: MEDLINE
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