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Trapping mammalian protein complexes in viral particles

Sven Eyckerman (UGent) , Kevin Titeca (UGent) , Emmy Van Quickelberghe (UGent) , Eva Cloots (UGent) , Annick Verhee (UGent) , Noortje Samyn (UGent) , Leentje De Ceuninck (UGent) , Evy Timmerman (UGent) , Delphine De Sutter (UGent) , Sam Lievens (UGent) , et al.
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Abstract
Cell lysis is an inevitable step in classical mass spectrometry-based strategies to analyse protein complexes. Complementary lysis conditions, in situ cross-linking strategies and proximal labelling techniques are currently used to reduce lysis effects on the protein complex. We have developed Virotrap, a viral particle sorting approach that obviates the need for cell homogenization and preserves the protein complexes during purification. By fusing a bait protein to the HIV-1 GAG protein, we show that interaction partners become trapped within virus-like particles (VLPs) that bud from mammalian cells. Using an efficient VLP enrichment protocol, Virotrap allows the detection of known binary interactions and MS-based identification of novel protein partners as well. In addition, we show the identification of stimulus-dependent interactions and demonstrate trapping of protein partners for small molecules. Virotrap constitutes an elegant complementary approach to the arsenal of methods to study protein complexes.
Keywords
MASS-SPECTROMETRY, TIR-DOMAIN, CELLS, HIV-1, UBIQUITINATION, DIMERIZATION, RELEASE, BINDING, MASPIT, MAPPIT

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Chicago
Eyckerman, Sven, Kevin Titeca, Emmy Van Quickelberghe, Eva Cloots, Annick Verhee, Noortje Samyn, Leentje De Ceuninck, et al. 2016. “Trapping Mammalian Protein Complexes in Viral Particles.” Nature Communications 7.
APA
Eyckerman, S., Titeca, K., Van Quickelberghe, E., Cloots, E., Verhee, A., Samyn, N., De Ceuninck, L., et al. (2016). Trapping mammalian protein complexes in viral particles. NATURE COMMUNICATIONS, 7.
Vancouver
1.
Eyckerman S, Titeca K, Van Quickelberghe E, Cloots E, Verhee A, Samyn N, et al. Trapping mammalian protein complexes in viral particles. NATURE COMMUNICATIONS. 2016;7.
MLA
Eyckerman, Sven, Kevin Titeca, Emmy Van Quickelberghe, et al. “Trapping Mammalian Protein Complexes in Viral Particles.” NATURE COMMUNICATIONS 7 (2016): n. pag. Print.
@article{8508990,
  abstract     = {Cell lysis is an inevitable step in classical mass spectrometry-based strategies to analyse protein complexes. Complementary lysis conditions, in situ cross-linking strategies and proximal labelling techniques are currently used to reduce lysis effects on the protein complex. We have developed Virotrap, a viral particle sorting approach that obviates the need for cell homogenization and preserves the protein complexes during purification. By fusing a bait protein to the HIV-1 GAG protein, we show that interaction partners become trapped within virus-like particles (VLPs) that bud from mammalian cells. Using an efficient VLP enrichment protocol, Virotrap allows the detection of known binary interactions and MS-based identification of novel protein partners as well. In addition, we show the identification of stimulus-dependent interactions and demonstrate trapping of protein partners for small molecules. Virotrap constitutes an elegant complementary approach to the arsenal of methods to study protein complexes.},
  articleno    = {11416},
  author       = {Eyckerman, Sven and Titeca, Kevin and Van Quickelberghe, Emmy and Cloots, Eva and Verhee, Annick and Samyn, Noortje and De Ceuninck, Leentje and Timmerman, Evy and De Sutter, Delphine and Lievens, Sam and Van Calenbergh, Serge and Gevaert, Kris and Tavernier, Jan},
  issn         = {2041-1723},
  journal      = {NATURE COMMUNICATIONS},
  language     = {eng},
  pages        = {8},
  title        = {Trapping mammalian protein complexes in viral particles},
  url          = {http://dx.doi.org/10.1038/ncomms11416},
  volume       = {7},
  year         = {2016},
}

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