RESEARCH ARTICLE


A Cell-Based Protein-Protein Interaction Method Using a Permuted Luciferase Reporter



Haifeng Eishingdrelo*, 1, Jidong Cai 2, Paul Weissensee 2, Praveen Sharma 2, Michael J Tocci 2, Paul S Wright 3
1 BioInvenu Corporation, USA, East Hanover, NJ
2 Sanofi R&D, Bridgewater, NJ
3 Sanofi R&D, Oro Valley, AZ, USA


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© Eishingdrelo et al.; Licensee Bentham Open.

open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

* Address correspondence to this author at the BioInvenu Corporation, USA, East Hanover, NJ, USA; Tel: 973-585-6777; Fax: 973-585-6776; E-mail: haifeng.eishingdrelo@bioinvenu.com


Abstract

We have developed a novel cell-based protein-protein interaction assay method. The method relies on conversion of an inactive permuted luciferase containing a Tobacco Etch Virus protease (TEV) cleavage sequence fused onto protein (A) to an active luciferase upon interaction and cleavage by another protein (B) fused with the TEV protease. We demonstrate assay applicability for ligand-induced protein-protein interactions including G-protein coupled receptors, receptor tyrosine kinases and nuclear hormone receptors.

Keywords: Cell-based assay, permuted luciferase, protein-protein interactions, GPCR, NHR, RTK, cell signaling, beta-arrestins.