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

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).

SUBMITTER: Peng L

PROVIDER: S-EPMC9168144 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature

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

Peng Liyuan L Jiang Jingwen J Zhou Li L Nice Edouard C EC Huang Canhua C

STAR protocols 20220604 2

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 d ...[more]

PMID: 35677609

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