title: A phage display system designed to detect and study protein-protein interactions.
abstract: Analysing protein-protein interactions is critical in proteomics and drug discovery. The usage of 2-Hybrid (2lambda) systems is limited to an in vivo environment. We describe a bacteriophage 2-Hybrid system for studying protein interactions in vitro. Bait and prey are displayed as fusions to the surface of phage lambda that are marked with different selectable drug-resistant markers. An interaction of phages in vitro through displayed proteins allows bacterial infection by two phages resulting in double drug-resistant bacterial colonies at very low multiplicity of infections. We demonstrate interaction of the protein sorting signal Ubiquitin with the Vps9-CUE, a Ubiquitin binding domain, and by the interaction of (Gly-Glu)(4) and (Gly-Arg)(4) peptides. Interruptions of the phage interactions by non-fused (free) bait or prey molecules show how robust and unique our approach is. We also demonstrate the use of Ubiquitin and CUE display phages to find binding partners in a lambda-display library. The unique usefulness to 2lambda is also described.
authors.affiliation: Laboratory of Molecular Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Catherine L
authors.initials: CL
authors.lastname: Bair
authors.forename: Amos
authors.initials: A
authors.lastname: Oppenheim
authors.forename: Andrei
authors.initials: A
authors.lastname: Trostel
authors.forename: Gali
authors.initials: G
authors.lastname: Prag
authors.forename: Sankar
authors.initials: S
authors.lastname: Adhya
mesh_terms: D010582:Bacteriophage lambda / Q000378:metabolism*; D005821:Genetic Techniques; D005822:Genetic Vectors; D009842:Oligopeptides / Q000378:metabolism; D019151:Peptide Library; D010957:Plasmids; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D040901:Proteomics; D020798:Two-Hybrid System Techniques; D025801:Ubiquitin / Q000378:metabolism*
chemical_list: D009842:Oligopeptides; D019151:Peptide Library; D011506:Proteins; D025801:Ubiquitin