2. Academic Validation
  3. Trapping mammalian protein complexes in viral particles

Trapping mammalian protein complexes in viral particles

  • Nat Commun. 2016 Apr 28:7:11416. doi: 10.1038/ncomms11416.
Sven Eyckerman 1 2,  Kevin Titeca 1 2,  Emmy Van Quickelberghe 1 2,  Eva Cloots 1 2,  Annick Verhee 1 2,  Noortje Samyn 1 2,  Leentje De Ceuninck 1 2,  Evy Timmerman 1 2,  Delphine De Sutter 1 2,  Sam Lievens 1 2,  Serge Van Calenbergh 3,  Kris Gevaert 1 2,  Jan Tavernier 1 2
Affiliations

Affiliations

  • 1 VIB Medical Biotechnology Center, VIB, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.
  • 2 Department of Biochemistry, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.
  • 3 Laboratory for Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Ghent University, Harelbekestraat 72, Ghent B-9000, Belgium.
PMID: 27122307 DOI: 10.1038/ncomms11416
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.

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