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Affinities of protein-protein interactions from single-cell titration experiments

Laboratory of Biology and Modelling of the Cell - ENS Lyon, France

mardi 11 décembre 2018 - 11h00 - Salle de conférence E1031, CBI
Invité(e) par Biologie structurale intégrative, 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.

Imprimer Envoyer

Université de Strasbourg

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