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A time-resolved fluorescence resonance energy transfer screening assay for discovery of protein-protein interaction modulators
Protein-protein interactions (PPIs) have emerged as promising yet challenging therapeutic targets. A robust bioassay is required for rapid PPI modulator discovery. Here, we present a time-resolved Förster's (fluorescence) resonance energy transfer assay protocol for PPI modulator screening in a...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8433285/ https://www.ncbi.nlm.nih.gov/pubmed/34527960 http://dx.doi.org/10.1016/j.xpro.2021.100804 |
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author | Tang, Cong Niu, Qiankun Cicka, Danielle Du, Yuhong Mo, Xiulei Fu, Haian |
author_facet | Tang, Cong Niu, Qiankun Cicka, Danielle Du, Yuhong Mo, Xiulei Fu, Haian |
author_sort | Tang, Cong |
collection | PubMed |
description | Protein-protein interactions (PPIs) have emerged as promising yet challenging therapeutic targets. A robust bioassay is required for rapid PPI modulator discovery. Here, we present a time-resolved Förster's (fluorescence) resonance energy transfer assay protocol for PPI modulator screening in a 1536-well plate format. We use hypomorph SMAD4(R361H)-SMAD3 PPI as an example to illustrate the application of the protocol for screening of variant-directed PPI inducers. This platform can be readily adapted for the discovery of both small-molecule PPI inducers and inhibitors. For complete details on the use and execution of this protocol, please refer to Tang et al. (2020). |
format | Online Article Text |
id | pubmed-8433285 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-84332852021-09-14 A time-resolved fluorescence resonance energy transfer screening assay for discovery of protein-protein interaction modulators Tang, Cong Niu, Qiankun Cicka, Danielle Du, Yuhong Mo, Xiulei Fu, Haian STAR Protoc Protocol Protein-protein interactions (PPIs) have emerged as promising yet challenging therapeutic targets. A robust bioassay is required for rapid PPI modulator discovery. Here, we present a time-resolved Förster's (fluorescence) resonance energy transfer assay protocol for PPI modulator screening in a 1536-well plate format. We use hypomorph SMAD4(R361H)-SMAD3 PPI as an example to illustrate the application of the protocol for screening of variant-directed PPI inducers. This platform can be readily adapted for the discovery of both small-molecule PPI inducers and inhibitors. For complete details on the use and execution of this protocol, please refer to Tang et al. (2020). Elsevier 2021-09-08 /pmc/articles/PMC8433285/ /pubmed/34527960 http://dx.doi.org/10.1016/j.xpro.2021.100804 Text en © 2021. 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 Tang, Cong Niu, Qiankun Cicka, Danielle Du, Yuhong Mo, Xiulei Fu, Haian A time-resolved fluorescence resonance energy transfer screening assay for discovery of protein-protein interaction modulators |
title | A time-resolved fluorescence resonance energy transfer screening assay for discovery of protein-protein interaction modulators |
title_full | A time-resolved fluorescence resonance energy transfer screening assay for discovery of protein-protein interaction modulators |
title_fullStr | A time-resolved fluorescence resonance energy transfer screening assay for discovery of protein-protein interaction modulators |
title_full_unstemmed | A time-resolved fluorescence resonance energy transfer screening assay for discovery of protein-protein interaction modulators |
title_short | A time-resolved fluorescence resonance energy transfer screening assay for discovery of protein-protein interaction modulators |
title_sort | time-resolved fluorescence resonance energy transfer screening assay for discovery of protein-protein interaction modulators |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8433285/ https://www.ncbi.nlm.nih.gov/pubmed/34527960 http://dx.doi.org/10.1016/j.xpro.2021.100804 |
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