A phage-based system to select multiple protein-protein interactions simultaneously from combinatorial libraries

• Published in: FEBS letters Vol. 440; no. 1; pp. 135 - 140
• Main Authors: Rudert, Fritz, Woltering, Claudia, Frisch, Christian, Rottenberger, Christine, Ilag, Leodevico L
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 27.11.1998
• Subjects: Adaptor Proteins, Vesicular Transport; Bacteriophages - genetics; Cell Cycle Proteins; Cloning, Molecular - methods; Combinatorial expression library; Fungal Proteins - genetics; Fungal Proteins - metabolism; Gene Library; Genetic Vectors; IR, interference-resistant; ISIS, in situ infectivity screening; Leucine Zippers; Membrane Proteins - genetics; Membrane Proteins - metabolism; Neurotrophin receptor; p75-ICD, p75 neurotrophin receptor intracellular domain; Peptide; Peptide Library; Phage display; Protein Binding; Protein-protein interaction; Proto-Oncogene Proteins c-jun - genetics; Proto-Oncogene Proteins c-jun - metabolism; Receptor, Nerve Growth Factor; Receptors, Nerve Growth Factor - genetics; Receptors, Nerve Growth Factor - metabolism; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Saccharomyces cerevisiae Proteins; Selectively infective phage; SIP, selectively infective phage; Transduction, Genetic; Viral Proteins - genetics; Viral Proteins - metabolism; Virus Assembly; Phage display; ISIS, in situ infectivity screening; p75-ICD, p75 neurotrophin receptor intracellular domain; Selectively infective phage; Protein-protein interaction; IR, interference-resistant; Neurotrophin receptor; SIP, selectively infective phage; Peptide; Combinatorial expression library
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(98)01413-6

Selectively infective phage (SIP) can be used to identify protein-protein interactions. SIP was modified to facilitate the simultaneous selection of interacting protein pairs from large combinatorial libraries. An interference-resistant phage was constructed which non-covalently, but stably links the genetic information of an interacting pair, encoded separately on phage and phagemid vectors, by co-packaging into hetero-polyphages. In a model system, the interaction between a SIP-selected peptide and the intracellular domain of the p75 neurotrophin receptor was detected in the presence of a 10 4-fold excess of a non-interacting control pair (jun leucine zipper and p75 intracellular domain) via SIP hetero-polyphage transductants. To minimize the redundancy of transductants and to minimize possible ligand exchange generated in a solution-based SIP screening, a filter-based in situ infectivity screening was developed. The combination of the above techniques may provide a powerful system for rapid screening of very large sequence spaces.