Aptamer-assisted two-point immobilized agonist-bound angiotensin II type 1 receptor for a second-site modulator discovery

• DOI: 10.1016/j.isci.2022.105361
• Publication Date: 2022-10-14T12:17:47Z
• Authors: Jiajun Liu; Ting Li; Ge Wang; Jiahuan Chen; Qingqing Yao; Qian Li; Xinfeng Zhao
• Keywords: Aptamer; Allosteric modulator

Methods of immobilized proteins are challenged by the way how to capture in their intact functional states. Here we present a two-point, high-specific method for immobilization conformationally specific angiotensin II type 1 receptor (AT1R) on amino-functionalized polystyrene microspheres. We identified selective DNA aptamer AT1R column-based SELEX approach with micromolar affinity. Two single-stranded strands were utilized introduce and 3-8 peptide microsphere surface, resulting two surface-positioned sites. The two-point exhibited enhanced ligand-binding activity stability comparison that prepared one-positioned site. Ginsenoside Rg1 rosmarinic acid screened from herbal extract proved bind through allosteric orthosteric sites receptor, respectively. These provide generally applicable protein conformation stability, ligand binding activity, screening efficiency.