Cited by 440Advanced Proteomics Techniques and ApplicationsBiotin and Related StudiesBioinformatics and Genomic Networks

Quantitative proteomics combined with BAC TransgeneOmics reveals in vivo protein interactions

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Hubner, Nina C., Bird, Alexander W., Cox, Jürgen, Splettstoesser, Bianca, Bandilla, Peter, Poser, Ina, Hyman, Anthony, Mann, Matthias·2010·Journal of Cell Biology

Abstract

Protein interactions are involved in all cellular processes. Their efficient and reliable characterization is therefore essential for understanding biological mechanisms. In this study, we show that combining bacterial artificial chromosome (BAC) TransgeneOmics with quantitative interaction proteomics, which we call quantitative BAC-green fluorescent protein interactomics (QUBIC), allows specific and highly sensitive detection of interactions using rapid, generic, and quantitative procedures with minimal material. We applied this approach to identify known and novel components of well-studied complexes such as the anaphase-promoting complex. Furthermore, we demonstrate second generation interaction proteomics by incorporating directed mutational transgene modification and drug perturbation into QUBIC. These methods identified domain/isoform-specific interactors of pericentrin- and phosphorylation-specific interactors of TACC3, which are necessary for its recruitment to mitotic spindles. The scalability, simplicity, cost effectiveness, and sensitivity of this method provide a basis for its general use in small-scale experiments and in mapping the human protein interactome.

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Authors
Hubner, N. C., Bird, A. W., Cox, J., Splettstoesser, B., Bandilla, P., Poser, I., Hyman, A., & Mann, M.
Affiliations
  • Max Planck Institute of Biochemistry (DE)
  • Max Planck Institute of Molecular Cell Biology and Genetics (DE)
Corresponding author
  • Hyman, Anthony
  • Mann, Matthias
Venue
Journal of Cell Biology
Published
2010-05-17
Volume
189
Issue
4
Pages
739-754
ISSN
  • 0021-9525 (ISSN-L)
  • 1540-8140
OpenAlex
W2059041864
Cited by
440
References
67
Citations by year
2026: 6 · 2025: 11 · 2024: 22 · 2023: 11 · 2022: 22 · 2021: 28 · 2020: 29 · 2019: 24 · 2018: 32 · 2017: 26 · 2016: 38 · 2015: 35 · 2014: 30 · 2013: 49 · 2012: 40
FWCI
18.51
Type
journal article
Language
en
Open access
Licence
cc-by-nc-sa
Locations
  • The Journal of Cell Biology (publishedVersion, cc-by-nc-sa)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-sa)
  • PubMed Central (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
  • Max Planck Institute for Plasma Physics (submittedVersion)
  • Max Planck Digital Library (submittedVersion)
  • MPG.PuRe (Max Planck Society) (submittedVersion)
Topics
  • Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Biotin and Related Studies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bioinformatics and Genomic Networks
Keywords
protein-protein interaction, QUBIC, anaphase-promoting complex, pericentrin, TACC3, mitotic spindle, drug-induced perturbation, protein isoforms
MeSH terms
Antigens, Fluorescent Antibody Technique, Humans, Models, Biological, Phosphorylation, Transgenes, Chromosomes, Artificial, Bacterial, Genomics, Protein Interaction Mapping, Proteomics

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