Trapping mammalian protein complexes in viral particles

Sven Eyckerman (), Kevin Titeca, Emmy Van Quickelberghe, Eva Cloots, Annick Verhee, Noortje Samyn, Leentje De Ceuninck, Evy Timmerman, Delphine De Sutter, Sam Lievens, Serge Van Calenbergh, Kris Gevaert and Jan Tavernier
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Sven Eyckerman: VIB Medical Biotechnology Center, VIB, Ghent University
Kevin Titeca: VIB Medical Biotechnology Center, VIB, Ghent University
Emmy Van Quickelberghe: VIB Medical Biotechnology Center, VIB, Ghent University
Eva Cloots: VIB Medical Biotechnology Center, VIB, Ghent University
Annick Verhee: VIB Medical Biotechnology Center, VIB, Ghent University
Noortje Samyn: VIB Medical Biotechnology Center, VIB, Ghent University
Leentje De Ceuninck: VIB Medical Biotechnology Center, VIB, Ghent University
Evy Timmerman: VIB Medical Biotechnology Center, VIB, Ghent University
Delphine De Sutter: VIB Medical Biotechnology Center, VIB, Ghent University
Sam Lievens: VIB Medical Biotechnology Center, VIB, Ghent University
Serge Van Calenbergh: Laboratory for Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Ghent University
Kris Gevaert: VIB Medical Biotechnology Center, VIB, Ghent University
Jan Tavernier: VIB Medical Biotechnology Center, VIB, Ghent University

Nature Communications, 2016, vol. 7, issue 1, 1-8

Abstract: 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.

Date: 2016
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DOI: 10.1038/ncomms11416

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