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
Identifiers and Pagination:Year: 2011
First Page: 122
Last Page: 128
Publisher Id: CCGTM-5-122
Article History:Received Date: 19/5/2011
Revision Received Date: 17/8/2011
Acceptance Date: 18/8/2011
Electronic publication date: 30/11/2011
Collection year: 2011
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.
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.