학술논문
Rapid, optimized interactomic screening
Document Type
Report
Author
Source
Nature Methods. June 1, 2015, p553, 10 p.
Subject
Language
English
ISSN
1548-7091
Abstract
High-throughput DNA sequencing facilitates whole-genome characterization within weeks (1,2). Likewise, advances in mass spectrometry (MS) (3,4) are enabling cellular proteomes to be defined. However, we have yet to exhaustively map [...]
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.
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.