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: 0301 basic medicine; DIMERIZATION; Science; UBIQUITINATION; HIV Infections; gag Gene Products, Human Immunodeficiency Virus; Article; 03 medical and health sciences; MASPIT; BINDING; Protein Interaction Mapping; Medicine and Health Sciences; Animals; Humans; RELEASE; 2. Zero hunger; 0303 health sciences; Q; Virion; Proteins; MASS-SPECTROMETRY; TIR-DOMAIN; 3. Good health; CELLS; HIV-1; MAPPIT; Protein Binding

AbstractCell 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.