Interactomics 2.0: From partially mapped qualitative protein-protein interaction graphs to biophysically characterized interaction networks
Gergö Gógl (U. Nice Côte-d’Azur)
For decades, the study of the human protein-protein interaction networks, a.k.a. the interactome has been dominated by qualitative « discovery » methods. While these tools have provided invaluable maps of cellular organization, they often lack the biophysical resolution required to understand the true dynamics of the cell and often miss weak and transient interactions that make up the bulk fabric of molecular interaction networks. We recently developed new generation interactomic tools, such as the native holdup approach, that allows us to shift from asking who interacts with whom to quantifying how strong is the driving force of each interaction and under what conditions they form. Our approach offer a high-throughput solution for the biophysical characterization of large-scale systems that, unlike traditional approaches, allows the direct measurement of equilibrium dissociation constants at proteome scales within complex cellular lysates taking into consideration indirect interactions, post-translational modifications and other biological mechanisms. Most importantly, we can finally study weak and transient complexes that were hiding in the blind spot of conventional interactomics for decades. We will discuss the technical implementation of nHU on large scale, the challenges of scaling quantitative measurements, and how this « bottom-up » biophysical data provides a more predictive framework for understanding cellular signaling and disease-state perturbations.