Department of Biophysics and Chemical Biology, Seoul National University, Seoul 08826, Korea.
CRI Center for Chemical Proteomics, Department of Chemistry, Seoul National University, Seoul 08826, Korea.
Cell Chem Biol. 2021 Mar 18;28(3):394-423. doi: 10.1016/j.chembiol.2020.12.001. Epub 2020 Dec 22.
Phenotype-based screening has emerged as an alternative route for discovering new chemical entities toward first-in-class therapeutics. However, clarifying their mode of action has been a significant bottleneck for drug discovery. For target protein identification, conventionally bioactive small molecules are conjugated onto solid supports and then applied to isolate target proteins from whole proteome. This approach requires a high binding affinity between bioactive small molecules and their target proteins. Besides, the binding affinity can be significantly hampered after structural modifications of bioactive molecules with linkers. To overcome these limitations, two major strategies have recently been pursued: (1) the covalent conjugation between small molecules and target proteins using photoactivatable moieties or electrophiles, and (2) label-free target identification through monitoring target engagement by tracking the thermal, proteolytic, or chemical stability of target proteins. This review focuses on recent advancements in target identification from covalent capturing to label-free strategies.
基于表型的筛选已经成为发现新的化学实体作为首创疗法的一种替代途径。然而，阐明其作用模式一直是药物发现的一个重大瓶颈。对于靶蛋白鉴定，传统上生物活性小分子与固体载体偶联，然后用于从整个蛋白质组中分离靶蛋白。这种方法需要生物活性小分子与其靶蛋白之间具有高结合亲和力。此外，生物活性分子与接头进行结构修饰后，结合亲和力会受到显著阻碍。为了克服这些限制，最近已经采用了两种主要策略：（1）使用光活化基团或亲电试剂，在小分子和靶蛋白之间进行共价偶联，（2）通过跟踪靶蛋白的热稳定性、蛋白水解稳定性或化学稳定性，来监测靶蛋白的结合，从而实现无标记的靶蛋白鉴定。本综述重点介绍了从共价捕获到无标记策略的靶蛋白鉴定的最新进展。
