Abstract
A new in vitro screening method has been developed and applied to a commercial phage-displayed cDNA library to search for novel protein-protein interactions. PDZ, WW and SH3 domains from PSD95, Nedd4, Src, Abl and Crk proteins were used as targets. 12 novel putative and 2 previously reported interactions were identified in test screens. The novel screening format, dubbed TAIS (target-assisted iterative screening), is discussed as an alternative platform to existing technologies for a pair-wise characterization of protein-protein interactions.
MeSH terms
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Amino Acid Sequence
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Calcium-Binding Proteins / metabolism*
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Chimerin Proteins / metabolism
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Disks Large Homolog 4 Protein
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Endosomal Sorting Complexes Required for Transport
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Humans
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Intracellular Signaling Peptides and Proteins
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Ligases / metabolism*
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Membrane Proteins / metabolism
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Molecular Sequence Data
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Myelin Proteins / metabolism
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NAV1.2 Voltage-Gated Sodium Channel
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Nedd4 Ubiquitin Protein Ligases
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Nerve Tissue Proteins / metabolism*
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Nogo Proteins
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Oligopeptides / metabolism*
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Protein Interaction Mapping
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Protein Structure, Tertiary
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Proto-Oncogene Proteins / metabolism*
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Proto-Oncogene Proteins c-abl / metabolism*
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Proto-Oncogene Proteins c-crk
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Recombinant Fusion Proteins / metabolism
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Sequence Homology
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Sodium Channels / metabolism
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Ubiquitin-Protein Ligases*
Substances
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Calcium-Binding Proteins
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Chimerin Proteins
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DLG4 protein, human
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Disks Large Homolog 4 Protein
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Endosomal Sorting Complexes Required for Transport
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Intracellular Signaling Peptides and Proteins
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Membrane Proteins
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Myelin Proteins
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NAV1.2 Voltage-Gated Sodium Channel
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Nerve Tissue Proteins
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Nogo Proteins
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Oligopeptides
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Proto-Oncogene Proteins
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Proto-Oncogene Proteins c-crk
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RTN4 protein, human
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Recombinant Fusion Proteins
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SCN2A protein, human
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Sodium Channels
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Src peptide
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postsynaptic density proteins
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LAPTM5 protein, human
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Nedd4 Ubiquitin Protein Ligases
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Nedd4 protein, human
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Ubiquitin-Protein Ligases
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Proto-Oncogene Proteins c-abl
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Ligases