Rapid, optimized interactomic screeningHakhverdyan, Z., Domanski, M., Hough, L. E., Oroskar, A. A., Oroskar, A. R., Keegan, S., Dilworth, D. J., Molloy, K. R., Sherman, V., Aitchison, J. D., Fenyoe, D., Chait, B. T., Jensen, T. H., Rout, M. P. & LaCava, J., 4-May-2015, In : Nature Methods. 12, 6, p. 553-560 10 p.
Research output: Contribution to journal › Article › Academic › peer-review
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.
|Number of pages||10|
|Publication status||Published - 4-May-2015|
- MASS-SPECTROMETRY, AFFINITY PURIFICATION, PROTEIN COMPLEXES, YEAST EXOSOME, MULTIPROTEIN COMPLEXES, ACTIN CYTOSKELETON, ARP2/3 COMPLEX, RNA-POLYMERASE, ORGANIZATION, NETWORK