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  3. A cellular thermal shift assay for detecting amino acid sites involved in drug target engagement

A cellular thermal shift assay for detecting amino acid sites involved in drug target engagement

  • STAR Protoc. 2022 Jun 4;3(2):101423. doi: 10.1016/j.xpro.2022.101423.
Liyuan Peng 1,  Jingwen Jiang 1,  Li Zhou 1,  Edouard C Nice 2,  Canhua Huang 1
Affiliations

Affiliations

  • 1 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.
  • 2 Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC 3800, Australia.
PMID: 35677609 DOI: 10.1016/j.xpro.2022.101423
Abstract

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 in situ. For complete details on the use and execution of this protocol, please refer to Peng et al. (2021).

Keywords

Cell Biology; Cell culture; Cell-based Assays; Protein Biochemistry; Protein expression and purification.

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