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
  • 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.