A cellular thermal shift assay for detecting amino acid sites involved in drug target engagement
Liyuan Peng, Jingwen Jiang, Li Zhou, Edouard C. Nice, Canhua Huang
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).
Topics & Concepts
DrugAmino acidDrug targetComputational biologyDrug developmentChemistryLigand (biochemistry)Drug discoveryBiochemistryCell biologyBiologyPharmacologyReceptorComputational Drug Discovery MethodsReceptor Mechanisms and SignalingCell Image Analysis Techniques