Trapping mammalian protein complexes in viral particles

• DOI: 10.1038/ncomms11416
• Publication Date: 2016-04-28T09:56:53Z
• Authors: Sven Eyckerman; Kevin Titeca; Emmy Van Quickelb...; Eva Cloots; Annick Verhee; Noortje Samyn; Leentje De Ceuninck; Evy Timmerman; Delphine De Sutter; Sam Lievens; Serge Van Calenbergh; Kris Gevaert; Jan Tavernier
• Keywords: Virus-like particle; Homogenization

Abstract Cell lysis is an inevitable step in classical mass spectrometry–based strategies to analyse protein complexes. Complementary conditions, situ cross-linking and proximal labelling techniques are currently used reduce effects on the complex. We have developed Virotrap, a viral particle sorting approach that obviates need for cell homogenization preserves complexes during purification. By fusing bait HIV-1 GAG protein, we show interaction partners become trapped within virus-like particles (VLPs) bud from mammalian cells. Using efficient VLP enrichment protocol, Virotrap allows detection of known binary interactions MS-based identification novel as well. In addition, stimulus-dependent demonstrate trapping small molecules. constitutes elegant complementary arsenal methods study