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

• Type of Material: Article
• Authors: Cong Tang, Emory University; Qiankun Niu, Emory University; Danielle Cicka, Emory University; Yuhong Du, Emory University; Xiulei Mo, Emory University; Haian Fu, Emory University
• Language: English
• Date: 2021-09-17
• Publisher: CellPress
• Publication Version: Final Published Version
• Copyright Statement: © 2021.
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1016/j.xpro.2021.100804
• Title of Journal or Parent Work: STAR Protocols
• Volume: 2
• Issue: 3
• Grant/Funding Information: This work was supported by the National Cancer Institute’s Office of Cancer Genomics Cancer Target Discovery and Development (CTD2) initiative (U01CA217875 to H.F.), the NCI Emory Lung Cancer SPORE (P50CA217691 to H.F.) Career Enhancement Program (P50CA217691 to X.M.), NCI MERIT Award (R37CA255459 to X.M.), Emory URC Award (to X.M.), the Imagine, Innovate and Impact (I3) Funds from the Emory School of Medicine, and through the Georgia CTSA NIH award (UL1-TR002378) and Winship Cancer Institute (NIH 5P30CA138292). C.T. was a visiting student in the Emory University School of Medicine- Xi'an Jiaotong University Health Science Center student exchange program and is a current recipient of the Marie Skłodowska-Curie grant (agreement No 101038053) from the European Union's Horizon 2020 research and innovation program.
• Keywords: High Throughput Screening; Cancer; Molecular/Chemical Probes
• Research Categories: Biology, Molecular; Health Sciences, Oncology

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 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.