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

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 sta...

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Detalles Bibliográficos
Autores principales: Peng, Liyuan, Jiang, Jingwen, Zhou, Li, Nice, Edouard C., Huang, Canhua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9168144/
https://www.ncbi.nlm.nih.gov/pubmed/35677609
http://dx.doi.org/10.1016/j.xpro.2022.101423
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author Peng, Liyuan
Jiang, Jingwen
Zhou, Li
Nice, Edouard C.
Huang, Canhua
author_facet Peng, Liyuan
Jiang, Jingwen
Zhou, Li
Nice, Edouard C.
Huang, Canhua
author_sort Peng, Liyuan
collection PubMed
description 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).
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spelling pubmed-91681442022-06-07 A cellular thermal shift assay for detecting amino acid sites involved in drug target engagement Peng, Liyuan Jiang, Jingwen Zhou, Li Nice, Edouard C. Huang, Canhua STAR Protoc Protocol 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). Elsevier 2022-06-04 /pmc/articles/PMC9168144/ /pubmed/35677609 http://dx.doi.org/10.1016/j.xpro.2022.101423 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Peng, Liyuan
Jiang, Jingwen
Zhou, Li
Nice, Edouard C.
Huang, Canhua
A cellular thermal shift assay for detecting amino acid sites involved in drug target engagement
title A cellular thermal shift assay for detecting amino acid sites involved in drug target engagement
title_full A cellular thermal shift assay for detecting amino acid sites involved in drug target engagement
title_fullStr A cellular thermal shift assay for detecting amino acid sites involved in drug target engagement
title_full_unstemmed A cellular thermal shift assay for detecting amino acid sites involved in drug target engagement
title_short A cellular thermal shift assay for detecting amino acid sites involved in drug target engagement
title_sort cellular thermal shift assay for detecting amino acid sites involved in drug target engagement
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9168144/
https://www.ncbi.nlm.nih.gov/pubmed/35677609
http://dx.doi.org/10.1016/j.xpro.2022.101423
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