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 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.
VIB Medical Biotechnology Center, VIB, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.|Department of Biochemistry, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.
Sven
S
Eyckerman
VIB Medical Biotechnology Center, VIB, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.|Department of Biochemistry, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.
Kevin
K
Titeca
VIB Medical Biotechnology Center, VIB, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.|Department of Biochemistry, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.
Emmy
E
Van Quickelberghe
VIB Medical Biotechnology Center, VIB, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.|Department of Biochemistry, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.
Eva
E
Cloots
VIB Medical Biotechnology Center, VIB, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.|Department of Biochemistry, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.
Annick
A
Verhee
VIB Medical Biotechnology Center, VIB, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.|Department of Biochemistry, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.
Noortje
N
Samyn
VIB Medical Biotechnology Center, VIB, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.|Department of Biochemistry, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.
Leentje
L
De Ceuninck
VIB Medical Biotechnology Center, VIB, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.|Department of Biochemistry, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.
Evy
E
Timmerman
VIB Medical Biotechnology Center, VIB, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.|Department of Biochemistry, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.
Delphine
D
De Sutter
VIB Medical Biotechnology Center, VIB, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.|Department of Biochemistry, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.
Sam
S
Lievens
Laboratory for Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Ghent University, Harelbekestraat 72, Ghent B-9000, Belgium.
Serge
S
Van Calenbergh
VIB Medical Biotechnology Center, VIB, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.|Department of Biochemistry, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.
Kris
K
Gevaert
VIB Medical Biotechnology Center, VIB, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.|Department of Biochemistry, Ghent University, A. Baertsoenkaai 3, Ghent B-9000, Belgium.
Jan
J
Tavernier
D000818:Animals; D015658:HIV Infections / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000235:genetics / Q000378:metabolism; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism*
D011506:Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus