Department of Urology, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University; The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen Urology Minimally Invasive Engineering Center, Shenzhen, Guangdong, China.
Department of Geriatrics, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University; The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen, Guangdong, China.
Med Res Rev. 2021 Nov;41(6):2893-2926. doi: 10.1002/med.21788. Epub 2021 Feb 3.
Small-molecule drugs modulate biological processes and disease states through engagement of target proteins in cells. Assessing drug-target engagement on a proteome-wide scale is of utmost importance in better understanding the molecular mechanisms of action of observed beneficial and adverse effects, as well as in developing next generation tool compounds and drugs with better efficacies and specificities. However, systematic assessment of drug-target engagement has been an arduous task. With the continuous development of mass spectrometry-based proteomics instruments and techniques, various chemical proteomics approaches for drug target deconvolution (i.e., the identification of molecular target for drugs) have emerged. Among these, the label-free target deconvolution approaches that do not involve the chemical modification of compounds of interest, have gained increased attention in the community. Here we provide an overview of the basic principles and recent biological applications of the most important label-free methods including the cellular thermal shift assay, pulse proteolysis, chemical denaturant and protein precipitation, stability of proteins from rates of oxidation, drug affinity responsive target stability, limited proteolysis, and solvent-induced protein precipitation. The state-of-the-art technical implications and future outlook for the label-free approaches are also discussed.
小分子药物通过与细胞内的靶蛋白结合来调节生物过程和疾病状态。在更好地理解观察到的有益和不利作用的分子作用机制，以及开发具有更好疗效和特异性的下一代工具化合物和药物方面，全面评估药物-靶标结合至关重要。然而，系统评估药物-靶标结合一直是一项艰巨的任务。随着基于质谱的蛋白质组学仪器和技术的不断发展，已经出现了各种用于药物靶标解析（即鉴定药物的分子靶标）的化学蛋白质组学方法。在这些方法中，不涉及感兴趣化合物的化学修饰的无标记靶标解析方法在该领域受到了越来越多的关注。本文概述了最重要的无标记方法的基本原理和最新生物学应用，包括细胞热转移分析、脉冲蛋白水解、化学变性剂和蛋白沉淀、氧化速率的蛋白稳定性、药物亲和反应靶标稳定性、有限蛋白水解和溶剂诱导蛋白沉淀。还讨论了无标记方法的最新技术意义和未来展望。
