A Split-Luciferase Reporter Recognizing GFP and mCherry Tags to Facilitate Studies of Protein–Protein Interactions
AbstractThe 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
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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.
Moustaqil M, Bhumkar A, Gonzalez L, Raoul L, Hunter DJB, Carrive P, Sierecki E, Gambin Y. A Split-Luciferase Reporter Recognizing GFP and mCherry Tags to Facilitate Studies of Protein–Protein Interactions. International Journal of Molecular Sciences. 2017; 18(12):2681.Chicago/Turabian Style
Moustaqil, Mehdi; Bhumkar, Akshay; Gonzalez, Laura; Raoul, Lisa; Hunter, Dominic J.B.; Carrive, Pascal; Sierecki, Emma; Gambin, Yann. 2017. "A Split-Luciferase Reporter Recognizing GFP and mCherry Tags to Facilitate Studies of Protein–Protein Interactions." Int. J. Mol. Sci. 18, no. 12: 2681.
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