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          Institute: MPI für molekulare Zellbiologie und Genetik     Collection: Publikationen MPI-CBG 2010-arch     Display Documents



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ID: 546684.0, MPI für molekulare Zellbiologie und Genetik / Publikationen MPI-CBG 2010-arch
Quantitative proteomics combined with BAC TransgeneOmics reveals in vivo protein interactions.
Authors:Hubner, Nina C.; Bird, Alexander W.; Cox, Jürgen; Splettstoesser, Bianca; Bandilla, Peter; Poser, Ina; Hyman, Anthony A.; Mann, Matthias
Date of Publication (YYYY-MM-DD):2010
Title of Journal:The Journal of Cell Biology
Volume:189
Issue / Number:4
Start Page:739
End Page:754
Copyright:not available
Audience:Experts Only
Intended Educational Use:No
Abstract / Description: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.
External Publication Status:published
Document Type:Article
Version Comment:Automatic journal name synchronization
Communicated by:nn
Affiliations:MPI für molekulare Zellbiologie und Genetik
Identifiers:LOCALID:4183
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