Diverse recognition of non-PxxP peptide ligands by the SH3 domains from p67(phox), Grb2 and Pex13p

EMBO J. 2002 Aug 15;21(16):4268-76. doi: 10.1093/emboj/cdf428.

Abstract

The basic function of the Src homology 3 (SH3) domain is considered to be binding to proline-rich sequences containing a PxxP motif. Recently, many SH3 domains, including those from Grb2 and Pex13p, were reported to bind sequences lacking a PxxP motif. We report here that the 22 residue peptide lacking a PxxP motif, derived from p47(phox), binds to the C-terminal SH3 domain from p67(phox). We applied the NMR cross-saturation method to locate the interaction sites for the non-PxxP peptides on their cognate SH3 domains from p67(phox), Grb2 and Pex13p. The binding site of the Grb2 SH3 partially overlapped the conventional PxxP-binding site, whereas those of p67(phox) and Pex13p SH3s are located in different surface regions. The non-PxxP peptide from p47(phox) binds to the p67(phox) SH3 more tightly when it extends to the N-terminus to include a typical PxxP motif, which enabled the structure determination of the complex, to reveal that the non-PxxP peptide segment interacted with the p67(phox) SH3 in a compact helix-turn-helix structure (PDB entry 1K4U).

MeSH terms

  • Adaptor Proteins, Signal Transducing*
  • Amino Acid Motifs
  • Amino Acid Sequence
  • Binding Sites
  • CSK Tyrosine-Protein Kinase
  • Escherichia coli
  • GRB2 Adaptor Protein
  • Humans
  • Ligands
  • Membrane Proteins / metabolism*
  • Models, Molecular
  • Molecular Sequence Data
  • Mutation
  • NADPH Oxidases
  • Peptides, Cyclic / metabolism
  • Phosphoproteins / metabolism*
  • Protein Binding
  • Protein Conformation
  • Protein-Tyrosine Kinases / metabolism
  • Proteins / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Saccharomyces cerevisiae Proteins / metabolism
  • Sequence Deletion
  • src Homology Domains / physiology*
  • src-Family Kinases

Substances

  • Adaptor Proteins, Signal Transducing
  • GRB2 Adaptor Protein
  • GRB2 protein, human
  • Ligands
  • Membrane Proteins
  • PEX13 protein, S cerevisiae
  • PEX13 protein, human
  • Peptides, Cyclic
  • Phosphoproteins
  • Proteins
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • cyclo-acetyl-(cysteinyl-histidyl-phenylalanyl-glutaminyl-phenylalanyl-cysteinyl)amide
  • neutrophil cytosol factor 67K
  • NADPH Oxidases
  • neutrophil cytosolic factor 1
  • Protein-Tyrosine Kinases
  • CSK Tyrosine-Protein Kinase
  • src-Family Kinases
  • CSK protein, human

Associated data

  • PDB/1K4U