Published: May 4, 2015
Main image from Hough et al. 2015 Nature Methods

Zhanna Hakhverdyan, Michal Domanski, Loren E Hough, Asha A Oroskar, Anil R Oroskar, Sarah Keegan, David J Dilworth, Kelly R Molloy, Vadim Sherman, John D Aitchison, David Fenyö, Brian T Chait, Torben Heick Jensen, Michael P Rout, John LaCava (2015). Nature Methods volume12, pages553–560. DOI: https://doi.org/10.1038/nmeth.3395

We must reliably map the interactomes of cellular macromolecular complexes in order to fully explore and understand biological systems. However, there are no methods to accurately predict how to capture a given macromolecular complex with its physiological binding partners. Here, we present a screening method that comprehensively explores the parameters affecting the stability of interactions in affinity-captured complexes, enabling the discovery of physiological binding partners in unparalleled detail. We have implemented this screen on several macromolecular complexes from a variety of organisms, revealing novel profiles for even well-studied proteins. Our approach is robust, economical and automatable, providing inroads to the rigorous, systematic dissection of cellular interactomes.