Trapping mammalian protein complexes in viral particles

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
DOI: 10.1038/ncomms11416 Publication Date: 2016-04-28T09:56:53Z
ABSTRACT
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.
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