| Title: |
A Split-Luciferase Reporter Recognizing GFP and mCherry Tags to Facilitate Studies of Protein–Protein Interactions |
| Authors: |
Mehdi Moustaqil; Akshay Bhumkar; Laura Gonzalez; Lisa Raoul; Dominic J. B. Hunter; Pascal Carrive; Emma Sierecki; Yann Gambin |
| Source: |
International Journal of Molecular Sciences, Vol 18, Iss 12, p 2681 (2017) |
| Publisher Information: |
MDPI AG |
| Publication Year: |
2017 |
| Collection: |
Directory of Open Access Journals: DOAJ Articles |
| Subject Terms: |
protein–protein interaction; split-luciferase; universal reporter; Leishmania tarentolae cell-free; Biology (General); QH301-705.5; Chemistry; QD1-999 |
| Description: |
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. |
| Document Type: |
article in journal/newspaper |
| Language: |
English |
| Relation: |
https://www.mdpi.com/1422-0067/18/12/2681; https://doaj.org/toc/1422-0067; https://doaj.org/article/67cea85b4efd4bf38d7b3901b180e724 |
| DOI: |
10.3390/ijms18122681 |
| Availability: |
https://doi.org/10.3390/ijms18122681; https://doaj.org/article/67cea85b4efd4bf38d7b3901b180e724 |
| Accession Number: |
edsbas.B2CCE08F |
| Database: |
BASE |