A phage-based system to select multiple protein-protein interactions simultaneously from combinatorial libraries.
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 heteropolyphages. 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.
Rudert
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MorphoSys AG, Martinsried/Munich, Germany. rudert@morphosys.de
Woltering
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Frisch
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Rottenberger
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Ilag
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D033942:Adaptor Proteins, Vesicular Transport; D001435:Bacteriophages / Q000235:genetics*; D018797:Cell Cycle Proteins; D003001:Cloning, Molecular / Q000379:methods*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015723:Gene Library; D005822:Genetic Vectors; D016350:Leucine Zippers; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D019151:Peptide Library; D011485:Protein Binding; D016755:Proto-Oncogene Proteins c-jun / Q000235:genetics / Q000378:metabolism; D020800:Receptor, Nerve Growth Factor; D017475:Receptors, Nerve Growth Factor / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D014161:Transduction, Genetic; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D019065:Virus Assembly
D033942:Adaptor Proteins, Vesicular Transport; D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D008565:Membrane Proteins; C071657:PEP3 protein, S cerevisiae; D019151:Peptide Library; D016755:Proto-Oncogene Proteins c-jun; D020800:Receptor, Nerve Growth Factor; D017475:Receptors, Nerve Growth Factor; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014764:Viral Proteins