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

Tang et al. (2021) STAR Protocols. CC BY-NC-ND 4.0

Tang et al. (2021) STAR Protocols. CC BY-NC-ND 4.0

Tang C, Niu Q, Cicka D, Du Y, Mo X, Fu H.

STAR Protocols

September 08, 2021

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 SMAD4R361H-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).

Last updated: October 04, 2021