biostudies-literatureHakhverdyan ZNovo Nordisk FondenNIGMS NIH HHS553-60https://www.ebi.ac.uk/biostudies/studies/S-EPMC4449307biostudies-literatureUnknown12(6)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.Nature methodsRapid, optimized interactomic screening.PMC4449307P50 GM076547U54 GM103511P41 GM103314NNF12OC0001211P41 GM109824Hough LEJensen THDilworth DJOroskar ARDomanski MRout MPMolloy KRHakhverdyan ZLaCava JChait BTSherman VAitchison JDKeegan SFenyo DOroskar AAfalseRapid, optimized interactomic screening.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.2015-01-01T00:00:00Z2015 Jun2025-04-18T16:38:44.617Z2019-03-27T01:52:26ZS-EPMC44493072593837010.1038/nmeth.3395