Du Yulin, Lyu Yifan, Li Shiquan, Ding Ding, Chen Jianghuai, Yang Cai, Sun Yang, Qu Fengli, Xiao Zeyu, Jiang Jianhui, Tan Weihong
Molecular Science and Biomedicine Laboratory (MBL), State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, Aptamer Engineering Center of Hunan Province, Hunan University, Changsha, Hunan 410082, China.
Shenzhen Research Institute, Hunan University, Shenzhen, Guangdong 518000, China.
Angew Chem Int Ed Engl. 2023 Mar 1;62(10):e202215387. doi: 10.1002/anie.202215387. Epub 2022 Dec 29.
Cell-specific aptamers offer a powerful tool to study membrane receptors at the single-molecule level. Most target receptors of aptamers are highly expressed on the cell surface, but difficult to analyze in situ because of dense distribution and fast velocity. Therefore, we herein propose a random sampling-based analysis strategy termed ligand dilution analysis (LDA) for easily implemented aptamer-based receptor study. Receptor density on the cell surface can be calculated based on a regression model. By using a synergistic ligand dilution design, colocalization and differentiation of aptamer and monoclonal antibody (mAb) binding on a single receptor can be realized. Once this is accomplished, precise binding site and detailed aptamer-receptor binding mode can be further determined using molecular docking and molecular dynamics simulation. The ligand dilution strategy also sets the stage for an aptamer-based dynamics analysis of two- and three-dimensional motion and fluctuation of highly expressed receptors on the live cell membrane.
细胞特异性适配体为在单分子水平研究膜受体提供了一个强大的工具。适配体的大多数靶受体在细胞表面高度表达，但由于分布密集和速度快，难以原位分析。因此，我们在此提出一种基于随机抽样的分析策略，称为配体稀释分析（LDA），用于轻松实现基于适配体的受体研究。细胞表面的受体密度可以基于回归模型进行计算。通过使用协同配体稀释设计，可以实现适配体与单克隆抗体（mAb）在单个受体上结合的共定位和区分。一旦完成此操作，可以使用分子对接和分子动力学模拟进一步确定精确的结合位点和详细的适配体-受体结合模式。配体稀释策略还为基于适配体的活细胞膜上高表达受体的二维和三维运动及波动的动力学分析奠定了基础。
