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Trapping mammalian protein complexes in viral particles

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 Virot...

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Detalles Bibliográficos
Autores principales: Eyckerman, Sven, Titeca, Kevin, Van Quickelberghe, Emmy, Cloots, Eva, Verhee, Annick, Samyn, Noortje, De Ceuninck, Leentje, Timmerman, Evy, De Sutter, Delphine, Lievens, Sam, Van Calenbergh, Serge, Gevaert, Kris, Tavernier, Jan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4853472/
https://www.ncbi.nlm.nih.gov/pubmed/27122307
http://dx.doi.org/10.1038/ncomms11416
Descripción
Sumario: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.