Mapping differential interactomes by affinity purification coupled with data independent mass spectrometry acquisition
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Author
Lambert, Jean-Philippe
Ivosev, Gordana
Couzens, Amber L.
Larsen, Brett
Taipale, Mikko
Lin, Zhen-Yuan
Zhong, Quan
Lindquist, Susan
Aebersold, Ruedi
Pawson, Tony
Bonner, Ron
Tate, Stephen
Gingras, Anne-Claude
Note: Order does not necessarily reflect citation order of authors.
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https://doi.org/10.1038/nmeth.2702Metadata
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Lambert, J., G. Ivosev, A. L. Couzens, B. Larsen, M. Taipale, Z. Lin, Q. Zhong, et al. 2013. “Mapping differential interactomes by affinity purification coupled with data independent mass spectrometry acquisition.” Nature methods 10 (12): 10.1038/nmeth.2702. doi:10.1038/nmeth.2702. http://dx.doi.org/10.1038/nmeth.2702.Abstract
Characterizing changes in protein-protein interactions associated with sequence variants (e.g. disease-associated mutations or splice forms) or following exposure to drugs, growth factors or hormones is critical to understanding how protein complexes are built, localized and regulated. Affinity purification (AP) coupled with mass spectrometry permits the analysis of protein interactions under near-physiological conditions, yet monitoring interaction changes requires the development of a robust and sensitive quantitative approach, especially for large-scale studies where cost and time are major considerations. To this end, we have coupled AP to data-independent mass spectrometric acquisition (SWATH), and implemented an automated data extraction and statistical analysis pipeline to score modulated interactions. Here, we use AP-SWATH to characterize changes in protein-protein interactions imparted by the HSP90 inhibitor NVP-AUY922 or melanoma-associated mutations in the human kinase CDK4. We show that AP-SWATH is a robust label-free approach to characterize such changes, and propose a scalable pipeline for systems biology studies.Other Sources
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3882083/pdf/Terms of Use
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