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Array MAPPIT: High-Throughput Interactome Analysis in Mammalian Cells

Sam Lievens (UGent) , Nele Vanderroost (UGent) , José Van Der Heyden (UGent) , Viola Gesellchen (UGent) , Marc Vidal and Jan Tavernier (UGent)
(2009) JOURNAL OF PROTEOME RESEARCH. 8(2). p.877-886
Author
Organization
Abstract
Physical interactions between proteins play a key role in probably every cellular process. Efforts to chart the protein interaction networks are ongoing in a number of model organisms using a diversity of approaches. The resulting genome-wide interaction maps will provide a scaffold for further detailed functional analysis. We developed MAPPIT, a mammalian two-hybrid approach that allows identification and analysis of mammalian protein-protein interactions in their native environment. Here, we introduce an efficient MAPPIT assay that permits high-throughput screening of arrayed collections of proteins and complements a previously published cDNA library screening approach. We validated both methods in screens for interaction partners of the Cullin-based E3 ubiquitin ligase subunits SKP1 and Elongin C. In addition to a number of known interactors, novel SKP1 and Elongin C binding proteins were identified. The array assay is an important addition to the MAPPIT suite of technologies that is expected to significantly increase its utility as a toolbox to screen for novel interactors of proteins or small molecules.
Keywords
YEAST, MAPPIT, MOTIFS, BIOLOGY, MOLECULAR-INTERACTIONS, screening, PROTEIN INTERACTION NETWORK, two-hybrid, COMPLEXES, protein-protein interaction, interactome, RESOURCE, LEPTIN RECEPTOR, TRAP, CIS

Citation

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MLA
Lievens, Sam, Nele Vanderroost, José Van Der Heyden, et al. “Array MAPPIT: High-Throughput Interactome Analysis in Mammalian Cells.” JOURNAL OF PROTEOME RESEARCH 8.2 (2009): 877–886. Print.
APA
Lievens, Sam, Vanderroost, N., Van Der Heyden, J., Gesellchen, V., Vidal, M., & Tavernier, J. (2009). Array MAPPIT: High-Throughput Interactome Analysis in Mammalian Cells. JOURNAL OF PROTEOME RESEARCH, 8(2), 877–886.
Chicago author-date
Lievens, Sam, Nele Vanderroost, José Van Der Heyden, Viola Gesellchen, Marc Vidal, and Jan Tavernier. 2009. “Array MAPPIT: High-Throughput Interactome Analysis in Mammalian Cells.” Journal of Proteome Research 8 (2): 877–886.
Chicago author-date (all authors)
Lievens, Sam, Nele Vanderroost, José Van Der Heyden, Viola Gesellchen, Marc Vidal, and Jan Tavernier. 2009. “Array MAPPIT: High-Throughput Interactome Analysis in Mammalian Cells.” Journal of Proteome Research 8 (2): 877–886.
Vancouver
1.
Lievens S, Vanderroost N, Van Der Heyden J, Gesellchen V, Vidal M, Tavernier J. Array MAPPIT: High-Throughput Interactome Analysis in Mammalian Cells. JOURNAL OF PROTEOME RESEARCH. WASHINGTON, DC 20036 USA: AMER CHEMICAL SOC; 2009;8(2):877–86.
IEEE
[1]
S. Lievens, N. Vanderroost, J. Van Der Heyden, V. Gesellchen, M. Vidal, and J. Tavernier, “Array MAPPIT: High-Throughput Interactome Analysis in Mammalian Cells,” JOURNAL OF PROTEOME RESEARCH, vol. 8, no. 2, pp. 877–886, 2009.
@article{539377,
  abstract     = {{Physical interactions between proteins play a key role in probably every cellular process. Efforts to chart the protein interaction networks are ongoing in a number of model organisms using a diversity of approaches. The resulting genome-wide interaction maps will provide a scaffold for further detailed functional analysis. We developed MAPPIT, a mammalian two-hybrid approach that allows identification and analysis of mammalian protein-protein interactions in their native environment. Here, we introduce an efficient MAPPIT assay that permits high-throughput screening of arrayed collections of proteins and complements a previously published cDNA library screening approach. We validated both methods in screens for interaction partners of the Cullin-based E3 ubiquitin ligase subunits SKP1 and Elongin C. In addition to a number of known interactors, novel SKP1 and Elongin C binding proteins were identified. The array assay is an important addition to the MAPPIT suite of technologies that is expected to significantly increase its utility as a toolbox to screen for novel interactors of proteins or small molecules.}},
  author       = {{Lievens, Sam and Vanderroost, Nele and Van Der Heyden, José and Gesellchen, Viola and Vidal, Marc and Tavernier, Jan}},
  issn         = {{1535-3893}},
  journal      = {{JOURNAL OF PROTEOME RESEARCH}},
  keywords     = {{YEAST,MAPPIT,MOTIFS,BIOLOGY,MOLECULAR-INTERACTIONS,screening,PROTEIN INTERACTION NETWORK,two-hybrid,COMPLEXES,protein-protein interaction,interactome,RESOURCE,LEPTIN RECEPTOR,TRAP,CIS}},
  language     = {{eng}},
  number       = {{2}},
  pages        = {{877--886}},
  publisher    = {{AMER CHEMICAL SOC}},
  title        = {{Array MAPPIT: High-Throughput Interactome Analysis in Mammalian Cells}},
  url          = {{http://dx.doi.org/10.1021/pr8005167}},
  volume       = {{8}},
  year         = {{2009}},
}

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