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

被引：3

作者：

Tang, Cong ^{[1
,2
,6
]}

Niu, Qiankun ^{[1
]}

Cicka, Danielle ^{[1
]}

Du, Yuhong ^{[1
,3
]}

Mo, Xiulei ^{[1
]}

Fu, Haian ^{[1
,3
,4
,5
]}

机构：

[1] Emory Univ, Sch Med, Dept Pharmacol & Chem Biol, Atlanta, GA 30322 USA

[2] Xi An Jiao Tong Univ, Affiliated Hosp 1, Med Sch, Xian 710061, Peoples R China

[3] Emory Univ, Sch Med, Emory Chem Biol Discovery Ctr, Atlanta, GA 30322 USA

[4] Emory Univ, Winship Canc Inst, Dept Hematol & Med Oncol, Atlanta, GA 30322 USA

[5] Emory Univ, Winship Canc Inst, Atlanta, GA 30322 USA

[6] Univ Lisbon, Fac Med, Inst Med Mol, Ave Prof Egas Moniz, P-1649028 Lisbon, Portugal

来源：

STAR PROTOCOLS | 2021年 / 2卷 / 03期

关键词：

Cancer; High Throughput Screening; Molecular/Chemical Probes;

D O I：

10.1016/j.xpro.2021.100804

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 Forster'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).

引用

页数：15

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