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A time-resolved fluorescence resonance energy transfer screening assay for discovery of protein-protein interaction modulators.

ABSTRACT: 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).

SUBMITTER: Tang C

PROVIDER: S-EPMC8433285 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature

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A time-resolved fluorescence resonance energy transfer screening assay for discovery of protein-protein interaction modulators.

Tang Cong C Niu Qiankun Q Cicka Danielle D Du Yuhong Y Mo Xiulei X Fu Haian H

STAR protocols 20210908 3

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<sup>R361H</sup>-SMAD3 PPI as an example to illustrate the application of the protocol for screening of variant-directed PPI inducers. This platform can be re ...[more]

PMID: 34527960

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