1. Trapping mammalian protein complexes in viral particles.
- Author
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Eyckerman S, Titeca K, Van Quickelberghe E, Cloots E, Verhee A, Samyn N, De Ceuninck L, Timmerman E, De Sutter D, Lievens S, Van Calenbergh S, Gevaert K, and Tavernier J
- Subjects
- Animals, HIV Infections genetics, HIV Infections virology, HIV-1 genetics, Humans, Protein Binding, Proteins genetics, Virion genetics, Virion metabolism, gag Gene Products, Human Immunodeficiency Virus genetics, HIV Infections metabolism, HIV-1 metabolism, Protein Interaction Mapping methods, Proteins metabolism, gag Gene Products, Human Immunodeficiency Virus metabolism
- 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.
- Published
- 2016
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