Improving analytical methods for protein-protein interaction through implementation of chemically inducible dimerization.

Details

Serval ID
serval:BIB_F1D921C90F97
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Improving analytical methods for protein-protein interaction through implementation of chemically inducible dimerization.
Journal
Scientific Reports
Author(s)
Andersen T.G., Nintemann S.J., Marek M., Halkier B.A., Schulz A., Burow M.
ISSN
2045-2322 (Electronic)
ISSN-L
2045-2322
Publication state
Published
Issued date
2016
Peer-reviewed
Oui
Volume
6
Pages
27766
Language
english
Notes
Publication types: Journal Article
Publication Status: epublish
Abstract
When investigating interactions between two proteins with complementary reporter tags in yeast two-hybrid or split GFP assays, it remains troublesome to discriminate true- from false-negative results and challenging to compare the level of interaction across experiments. This leads to decreased sensitivity and renders analysis of weak or transient interactions difficult to perform. In this work, we describe the development of reporters that can be chemically induced to dimerize independently of the investigated interactions and thus alleviate these issues. We incorporated our reporters into the widely used split ubiquitin-, bimolecular fluorescence complementation (BiFC)- and Förster resonance energy transfer (FRET)- based methods and investigated different protein-protein interactions in yeast and plants. We demonstrate the functionality of this concept by the analysis of weakly interacting proteins from specialized metabolism in the model plant Arabidopsis thaliana. Our results illustrate that chemically induced dimerization can function as a built-in control for split-based systems that is easily implemented and allows for direct evaluation of functionality.
Pubmed
Web of science
Open Access
Yes
Create date
01/07/2016 8:06
Last modification date
20/08/2019 17:19
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