Affinities of protein-protein interactions from single-cell titration experiments
Dr Martin SPICHTY
Laboratory of Biology and Modelling of the Cell - ENS Lyon, France
Tuesday, December 11th 2018 - 11 a.m.
- Conference room E1031, CBI
Hosted by Integrated structural Biology, Roland STOTE
I present a novel quantitative yeast two-hybrid system based on fluorescent fusion proteins. It allows simultaneous quantification of the reaction partners (Bait and Prey) and the reporter at the single-cell level by flow cytometry. After only two hours of reaction, expression of the reporter can easily be detected even for weakly interacting proteins (with dissociation constants in the micromolar range). In-cellula affinities of protein-protein interactions can be extracted from the acquired data through a titration-like analysis. As a proof of concept, we demonstrate the applicability of this system on a set of proteins from different families and organisms with dissociation constants ranging from picomolar to micromolar.