一种用于检测药物靶标结合涉及的氨基酸位点的细胞热转移分析。

State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University and Collaborative Innovation Center for Biotherapy, Chengdu 610041, P.R. China.

Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC 3800, Australia.

STAR Protoc. 2022 Jun 4;3(2):101423. doi: 10.1016/j.xpro.2022.101423. eCollection 2022 Jun 17.

Evaluating how drugs bind to specific targets at the cellular level is essential to their development and efficacy. Here, we present a protocol to detect the amino acid sites involved in drug-target engagement using a cellular thermal shift assay, which is based on ligand-induced protein thermal stabilization. We also describe generation of cell lines expressing wild-type or mutated target protein and drug treatment. This protocol enables a convenient way to assess amino acid sites involved in drug-target interaction . For complete details on the use and execution of this protocol, please refer to Peng et al. (2021).

评估药物在细胞水平上与特定靶点的结合方式对于它们的开发和疗效至关重要。在这里，我们提出了一种使用细胞热转移分析（CETSA）检测涉及药物-靶标结合的氨基酸位点的方案，该方法基于配体诱导的蛋白质热稳定性。我们还描述了表达野生型或突变靶蛋白的细胞系的生成和药物处理。该方案为评估药物-靶标相互作用涉及的氨基酸位点提供了一种方便的方法。有关该方案使用和执行的完整详细信息，请参阅 Peng 等人。（2021 年）。