Issue 3, 2014

A bidirectional fluorescent two-hybrid system for monitoring protein–protein interactions

Abstract

Two-hybrid systems have been the cornerstone of research into protein–protein interactions, but these systems typically rely on life/death reporters that put additional selective pressure on the host organism, and potentially lead to false positives. Here we report a bidirectional fluorescence-based bacterial two-hybrid system that enables both the association and dissociation of a given protein–protein interaction to be monitored. The functionality of this system and its compatibility with FACS screening are demonstrated in the forward and reverse direction using known interacting protein-partners and their cyclic peptide inhibitors. The reported fluorescent two-hybrid system may be used in the forward direction for the identification of interacting protein partners, or as a reverse two-hybrid system for the high-throughput identification of protein–protein interaction inhibitors.

Graphical abstract: A bidirectional fluorescent two-hybrid system for monitoring protein–protein interactions

Supplementary files

Article information

Article type
Paper
Submitted
30 Sep 2013
Accepted
29 Nov 2013
First published
29 Nov 2013
This article is Open Access
Creative Commons BY-NC license

Mol. BioSyst., 2014,10, 485-490

A bidirectional fluorescent two-hybrid system for monitoring protein–protein interactions

I. K. Nordgren and A. Tavassoli, Mol. BioSyst., 2014, 10, 485 DOI: 10.1039/C3MB70438F

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