Abstract
The GYF domain of CD2BP2 serves as an adapter that recognizes proline-rich sequences in intracellular proteins. Although the T cell adhesion molecule CD2 and the core splicing protein SmB/B' were previously shown to interact with CD2BP2-GYF, we are now using a general approach to identify putative GYF domain target sites within the human proteome. The phage display-derived recognition motif for CD2BP2-GYF is PPG(W/F/Y/M/L). SPOT analysis confirmed that the GYF domain interacts with peptides from human proteins containing the consensus site. Epitope mapping by NMR spectroscopy performed for several peptides revealed a conserved binding surface. A direct interaction of the CD2BP2-GYF domain with the novel protein interaction partners PI31 and NPWBP was verified by yeast two-hybrid analysis.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing
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Amino Acid Motifs
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Amino Acid Sequence
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Amino Acid Substitution
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Arginine / metabolism
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Carrier Proteins / chemistry*
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Carrier Proteins / genetics
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Carrier Proteins / metabolism*
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Chromatography, High Pressure Liquid
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Conserved Sequence
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Databases as Topic
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Glutathione Transferase / metabolism
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Humans
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Hydrophobic and Hydrophilic Interactions
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Lysine / metabolism
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Mass Spectrometry
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Molecular Sequence Data
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Nuclear Magnetic Resonance, Biomolecular
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Peptide Library
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Peptides / chemical synthesis
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Proline / chemistry
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Protein Structure, Secondary
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Protein Structure, Tertiary
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Proteome / analysis
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Recombinant Fusion Proteins / metabolism
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Trifluoroacetic Acid / pharmacology
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Two-Hybrid System Techniques
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Water / chemistry
Substances
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Adaptor Proteins, Signal Transducing
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CD2BP2 protein, human
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Carrier Proteins
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Peptide Library
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Peptides
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Proteome
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Recombinant Fusion Proteins
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Water
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Arginine
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Proline
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Trifluoroacetic Acid
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Glutathione Transferase
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Lysine