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Rapid, optimized interactomic screening.

ABSTRACT: 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.

SUBMITTER: Hakhverdyan Z

PROVIDER: S-EPMC4449307 | biostudies-other | 2015 Jun

REPOSITORIES: biostudies-other

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Rapid, optimized interactomic screening.

Hakhverdyan Zhanna Z Domanski Michal M Hough Loren E LE Oroskar Asha A AA Oroskar Anil R AR Keegan Sarah S Dilworth David J DJ Molloy Kelly R KR Sherman Vadim V Aitchison John D JD Fenyö David D Chait Brian T BT Jensen Torben Heick TH Rout Michael P MP LaCava John J

Nature methods 20150504 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 ...[more]

PMID: 25938370

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