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Unusual binding properties of the SH3 domain of the yeast actin-binding protein Abp1: structural and functional analysis.

Abp1p is an actin-binding protein that plays a central role in the organization of Saccharomyces cerevisiae actin cytoskeleton. By a combination of two-hybrid and phage-display approaches, we have identified six new ligands of the Abp1-SH3 domain. None of these SH3-mediated novel interactions was detected in recent all genome high throughput protein interaction projects. Here we show that the SH3-mediated association of Abp1p with the Ser/Thr kinases Prk1p and Ark1p is essential for their localization to actin cortical patches. The Abp1-SH3 domain has a rather unusual binding specificity, because its target peptides contain the tetrapentapeptide +XXXPXXPX+PXXL with positive charges flanking the polyproline core on both sides. Here we present the structure of the Abp1-SH3 domain solved at 1.3-A resolution. The peptide-binding pockets in the SH3 domain are flanked by two acidic residues that are uncommon at those positions in the SH3 domain family. We have shown by site-directed mutagenesis that one of these negatively charged side chains may be the key determinant for the preference for non-classical ligands.

Pubmed ID: 11668184


  • Fazi B
  • Cope MJ
  • Douangamath A
  • Ferracuti S
  • Schirwitz K
  • Zucconi A
  • Drubin DG
  • Wilmanns M
  • Cesareni G
  • Castagnoli L


The Journal of biological chemistry

Publication Data

February 15, 2002

Associated Grants

  • Agency: NIGMS NIH HHS, Id: GM42759
  • Agency: NIGMS NIH HHS, Id: GM50399

Mesh Terms

  • Actins
  • Amino Acid Motifs
  • Amino Acid Sequence
  • Binding Sites
  • Cytoskeleton
  • DNA-Binding Proteins
  • Endocytosis
  • Enzyme-Linked Immunosorbent Assay
  • Gene Library
  • Ligands
  • Models, Biological
  • Models, Molecular
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Peptide Library
  • Peptides
  • Plant Proteins
  • Plasmids
  • Protein Binding
  • Protein Conformation
  • Protein Structure, Tertiary
  • Receptor Protein-Tyrosine Kinases
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Schizosaccharomyces pombe Proteins
  • Structure-Activity Relationship
  • Transcription Factors
  • Two-Hybrid System Techniques
  • src Homology Domains