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Int. J. Mol. Sci. 2017, 18(12), 2681; doi:10.3390/ijms18122681

A Split-Luciferase Reporter Recognizing GFP and mCherry Tags to Facilitate Studies of Protein–Protein Interactions

1
European Molecular Biology Laboratory Australia (EMBL Australia) Node in Single Molecule Science, Sydney NSW 2031, Australia
2
School of Medical Sciences, The University of New South Wales, Sydney NSW 2031, Australia
These authors contributed equally.
Current address: Ecole Normale Superieure de Rennes, 35170 Bruz, France.
*
Authors to whom correspondence should be addressed.
Received: 31 October 2017 / Revised: 21 November 2017 / Accepted: 5 December 2017 / Published: 11 December 2017
(This article belongs to the Collection Proteins and Protein-Ligand Interactions)
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Abstract

The use of fluorescently-tagged proteins in microscopy has become routine, and anti-GFP (Green fluorescent protein) affinity matrices are increasingly used in proteomics protocols. However, some protein–protein interactions assays, such as protein complementation assays (PCA), require recloning of each protein as a fusion with the different parts of the complementation system. Here we describe a generic system where the complementation is separated from the proteins and can be directly used with fluorescently-tagged proteins. By using nanobodies and performing tests in cell-free expression systems, we accelerated the development of multiple reporters, detecting heterodimers and homodimers or oligomers tagged with GFP or mCherry. We demonstrate that the system can detect interactions at a broad range of concentrations, from low nanomolar up to micromolar. View Full-Text
Keywords: protein–protein interaction; split-luciferase; universal reporter; Leishmania tarentolae cell-free protein–protein interaction; split-luciferase; universal reporter; Leishmania tarentolae cell-free
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Moustaqil, M.; Bhumkar, A.; Gonzalez, L.; Raoul, L.; Hunter, D.J.B.; Carrive, P.; Sierecki, E.; Gambin, Y. A Split-Luciferase Reporter Recognizing GFP and mCherry Tags to Facilitate Studies of Protein–Protein Interactions. Int. J. Mol. Sci. 2017, 18, 2681.

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