Work

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

Year: 2021

Type: article

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) 29. Tang, C. ∙ Mo, X. ∙ Niu, Q. ... Hypomorph mutation-directed small-molecule protein-protein interaction inducers to restore mutant SMAD4-suppressed TGF-β signaling Cell Chem. Biol. 2020; 28 :636-647.e5 Google Scholar .

Source: STAR Protocols

DOI: 10.1016/j.xpro.2021.100804

Authors: Cong Tang, Qiankun Niu, Danielle Cicka, Yuhong Du, Xiulei Mo, Haian Fu

Institutions: Emory University, First Affiliated Hospital of Xi'an Jiaotong University, Xi'an Jiaotong University, Winship Cancer Institute

Language: English

Topic: Advanced Fluorescence Microscopy Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,500