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:

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.