Application of the Cellular Thermal Shift Assay (CETSA) to validate drug target engagement in platelets
Small molecule drugs play a major role in the study of human platelets. Effective action of a drug requires it to bind to one or more targets within the platelet (target engagement). However, although in vitro assays with isolated proteins can be used to determine drug affinity to these targets, add...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2024-12-01
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| Series: | Platelets |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/09537104.2024.2354833 |
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| author | Joanna-Marie Howes Matthew T. Harper |
| author_facet | Joanna-Marie Howes Matthew T. Harper |
| author_sort | Joanna-Marie Howes |
| collection | DOAJ |
| description | Small molecule drugs play a major role in the study of human platelets. Effective action of a drug requires it to bind to one or more targets within the platelet (target engagement). However, although in vitro assays with isolated proteins can be used to determine drug affinity to these targets, additional factors affect target engagement and its consequences in an intact platelet, including plasma membrane permeability, intracellular metabolism or compartmentalization, and level of target expression. Mechanistic interpretation of the effect of drugs on platelet activity requires comprehensive investigation of drug binding in the proper cellular context, i.e. in intact platelets. The Cellular Thermal Shift Assay (CETSA) is a valuable method to investigate target engagement within complex cellular environments. The assay is based on the principle that drug binding to a target protein increases that protein’s thermal stability. In this technical report, we describe the application of CETSA to platelets. We highlight CETSA as a quick and informative technique for confirming the direct binding of drugs to platelet protein targets, providing a platform for understanding the mechanism of action of drugs in platelets, and which will be a valuable tool for investigating platelet signaling and function. |
| format | Article |
| id | doaj-art-9aade0f0cb9a4384bad97a096502b8f7 |
| institution | OA Journals |
| issn | 0953-7104 1369-1635 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Platelets |
| spelling | doaj-art-9aade0f0cb9a4384bad97a096502b8f72025-08-20T02:12:19ZengTaylor & Francis GroupPlatelets0953-71041369-16352024-12-0135110.1080/09537104.2024.2354833Application of the Cellular Thermal Shift Assay (CETSA) to validate drug target engagement in plateletsJoanna-Marie Howes0Matthew T. Harper1Department of Pharmacology, University of Cambridge, Cambridge, UKDepartment of Pharmacology, University of Cambridge, Cambridge, UKSmall molecule drugs play a major role in the study of human platelets. Effective action of a drug requires it to bind to one or more targets within the platelet (target engagement). However, although in vitro assays with isolated proteins can be used to determine drug affinity to these targets, additional factors affect target engagement and its consequences in an intact platelet, including plasma membrane permeability, intracellular metabolism or compartmentalization, and level of target expression. Mechanistic interpretation of the effect of drugs on platelet activity requires comprehensive investigation of drug binding in the proper cellular context, i.e. in intact platelets. The Cellular Thermal Shift Assay (CETSA) is a valuable method to investigate target engagement within complex cellular environments. The assay is based on the principle that drug binding to a target protein increases that protein’s thermal stability. In this technical report, we describe the application of CETSA to platelets. We highlight CETSA as a quick and informative technique for confirming the direct binding of drugs to platelet protein targets, providing a platform for understanding the mechanism of action of drugs in platelets, and which will be a valuable tool for investigating platelet signaling and function.https://www.tandfonline.com/doi/10.1080/09537104.2024.2354833CETSAdrug bindingplateletsthermal stability |
| spellingShingle | Joanna-Marie Howes Matthew T. Harper Application of the Cellular Thermal Shift Assay (CETSA) to validate drug target engagement in platelets Platelets CETSA drug binding platelets thermal stability |
| title | Application of the Cellular Thermal Shift Assay (CETSA) to validate drug target engagement in platelets |
| title_full | Application of the Cellular Thermal Shift Assay (CETSA) to validate drug target engagement in platelets |
| title_fullStr | Application of the Cellular Thermal Shift Assay (CETSA) to validate drug target engagement in platelets |
| title_full_unstemmed | Application of the Cellular Thermal Shift Assay (CETSA) to validate drug target engagement in platelets |
| title_short | Application of the Cellular Thermal Shift Assay (CETSA) to validate drug target engagement in platelets |
| title_sort | application of the cellular thermal shift assay cetsa to validate drug target engagement in platelets |
| topic | CETSA drug binding platelets thermal stability |
| url | https://www.tandfonline.com/doi/10.1080/09537104.2024.2354833 |
| work_keys_str_mv | AT joannamariehowes applicationofthecellularthermalshiftassaycetsatovalidatedrugtargetengagementinplatelets AT matthewtharper applicationofthecellularthermalshiftassaycetsatovalidatedrugtargetengagementinplatelets |