School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, PR China.
School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, PR China.
J Chromatogr A. 2024 Aug 16;1729:465057. doi: 10.1016/j.chroma.2024.465057. Epub 2024 Jun 6.
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.
组胺 H1 受体（H1R）在过敏反应的发生中起着关键作用，因此需要设计一种基于 H1R 的高通量筛选方法，以有效地筛选新型配体。本研究提出了一种利用苯乙烯马来酸（SMA）提取和 His 标签共价键合来固定 H1R 膜蛋白的方法，最大限度地减少了非特异性蛋白质干扰，同时保持了天然蛋白质结构，并最大限度地提高了靶标暴露度。该方法用于开发高通量配体筛选的新型材料，并在细胞膜色谱（CMC）中实施。建立了 H1R-His-SMALPs/CMC 模型，并验证了其色谱性能（选择性、特异性和寿命），与之前的 CMC 模型相比，寿命有了显著提高。随后，该模型促进了化合物库中 H1R 配体的高通量筛选，并对潜在 H1R 拮抗剂的初步活性进行了验证。鉴定出一种新型的 H1R 拮抗剂为该领域的进一步发展奠定了基础。
