Titration of Apparent In-Cellula Affinities of Protein-Protein Interactions

Abstract

A genetic assay permits simultaneous quantification of two interacting proteins and their bound fraction at the single-cell level using flow cytometry. Apparent in-cellula affinities of protein-protein interactions can be extracted from the acquired data through a titration-like analysis. The applicability of this approach is demonstrated on a diverse set of interactions with proteins from different families and organisms and with in-vitro dissociation constants ranging from picomolar to micromolar.

Keywords: flow cytometry; protein-protein interactions; quantitative yeast-two hybrid; single-cell analysis.