Novel interaction partners of the CD2BP2-GYF domain.
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.
Kofler
Michael
M
Protein Engineering Group, Forschungsinstitut für Molekulare Pharmakologie and Freie Universität Berlin, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.
Motzny
Kathrin
K
Beyermann
Michael
M
Freund
Christian
C
D048868:Adaptor Proteins, Signal Transducing; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001120:Arginine / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D017124:Conserved Sequence; D019992:Databases as Topic; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008239:Lysine / Q000378:metabolism; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis; D011392:Proline / Q000737:chemistry; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D020543:Proteome / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014269:Trifluoroacetic Acid / Q000494:pharmacology; D020798:Two-Hybrid System Techniques; D014867:Water / Q000737:chemistry
D048868:Adaptor Proteins, Signal Transducing; C120378:CD2BP2 protein, human; D002352:Carrier Proteins; D019151:Peptide Library; D010455:Peptides; D020543:Proteome; D011993:Recombinant Fusion Proteins; D014867:Water; D001120:Arginine; D011392:Proline; D014269:Trifluoroacetic Acid; D005982:Glutathione Transferase; D008239:Lysine