Cited by 368Advanced Proteomics Techniques and ApplicationsBioinformatics and Genomic NetworksMass Spectrometry Techniques and Applications

Affinity-purification coupled to mass spectrometry: basic principles and strategies

Dunham, Wade H., Mullin, Michael, Gingras, Anne‐Claude·2012·PROTEOMICS

Abstract

Identifying the interactions established by a protein of interest can be a critical step in understanding its function. This is especially true when an unknown protein of interest is demonstrated to physically interact with proteins of known function. While many techniques have been developed to characterize protein-protein interactions, one strategy that has gained considerable momentum over the past decade for identification and quantification of protein-protein interactions, is affinity-purification followed by mass spectrometry (AP-MS). Here, we briefly review the basic principles used in affinity-purification coupled to mass spectrometry, with an emphasis on tools (both biochemical and computational), which enable the discovery and reporting of high quality protein-protein interactions.

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Crossref · 10.1002/pmic.201100523

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  • Limitations of the AP-MS experiment include the potential loss of transient or weakly bound complexes, the inherent noise associated with AP-MS relative to global proteomics analysis, and the short time point of the experiment, which precludes detection of degradation events with slow kinetics.

    “Limitations of this experiment include the potential loss of transient or weakly bound complexes in the enrichment and subsequent wash steps, the inherent noise associated with AP-MS relative to global proteomics analysis (Dunham et al., 2012; Yugandhar et al., 2019) and the short time point of the experiment, which precludes detection of degradation events with slow kinetics.”

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Authors
Dunham, W. H., Mullin, M., & Gingras, A.
Affiliations
Mount Sinai Hospital (CA)
Corresponding author
Gingras, Anne‐Claude
Venue
PROTEOMICS
Publisher
Wiley
Published
2012-05
Volume
12
Issue
10
Pages
1576-1590
ISSN
  • 1615-9853 (ISSN-L)
  • 1615-9861
OpenAlex
W1991953772
Cited by
368
References
144
Citations by year
2026: 16 · 2025: 33 · 2024: 29 · 2023: 24 · 2022: 27 · 2021: 33 · 2020: 24 · 2019: 31 · 2018: 27 · 2017: 37 · 2016: 19 · 2015: 23 · 2014: 27 · 2013: 13 · 2012: 4
FWCI
1.99
Type
journal article
Language
en
Open access
closed
Locations
  • PROTEOMICS (publishedVersion)
  • PubMed (publishedVersion)
Funders
Canadian Institutes of Health Research (MOP 84314)
Topics
  • Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bioinformatics and Genomic Networks
  • Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
Keywords
affinity purification, protein-protein interaction, affinity purification mass spectrometry, protein-protein interaction network, protein complex identification
MeSH terms
Animals, Chromatography, Affinity, Humans, Proteins, Mass Spectrometry, Protein Interaction Mapping, Systems Biology

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