A time-resolved fluorescence resonance energy transfer screening assay for discovery of protein-protein interaction modulators
- Resource Type
- article
- Authors
- Cong Tang; Qiankun Niu; Danielle Cicka; Yuhong Du; Xiulei Mo; Haian Fu
- Source
- STAR Protocols, Vol 2, Iss 3, Pp 100804- (2021)
- Subject
- Cancer
High Throughput Screening
Molecular/Chemical Probes
Science (General)
Q1-390
- Language
- English
- ISSN
- 2666-1667
Summary: 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).