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PubMed · 9862442

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

F Rudert·C Woltering·C Frisch·C Rottenberger·L L Ilag
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Abstract

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.

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Adaptor Proteins, Vesicular TransportBacteriophagesCell Cycle ProteinsCloning, MolecularFungal ProteinsGene LibraryGenetic VectorsLeucine ZippersMembrane ProteinsPeptide LibraryProtein BindingProto-Oncogene Proteins c-junReceptor, Nerve Growth FactorReceptors, Nerve Growth FactorRecombinant Fusion ProteinsSaccharomyces cerevisiae ProteinsTransduction, GeneticViral ProteinsVirus AssemblyJournal ArticleResearch Support, Non-U.S. Gov't

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F Rudert, C Woltering, C Frisch, C Rottenberger, L L Ilag. 1998-11-27. A phage-based system to select multiple protein-protein interactions simultaneously from combinatorial libraries.. https://doi.org/10.1016/s0014-5793(98)01413-6

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