Unusual binding properties of the SH3 domain of the yeast actin-binding protein Abp1: structural and functional analysis
| Autor: | Fazi, B, Cope, M, Douangamath, A, Ferracuti, S, Schirwitz, K, Zucconi, A, Drubin, D, Wilmanns, M, Cesareni, G, Castagnoli, L |
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| Rok vydání: | 2001 |
| Předmět: |
Models
Molecular Protein Structure Saccharomyces cerevisiae Proteins Protein Conformation Amino Acid Motifs Molecular Sequence Data Enzyme-Linked Immunosorbent Assay Saccharomyces cerevisiae Protein Serine-Threonine Kinases Ligands Models Biological src Homology Domains Structure-Activity Relationship Models Peptide Library Two-Hybrid System Techniques Site-Directed Amino Acid Sequence Cytoskeleton Gene Library Plant Proteins Binding Sites Molecular Receptor Protein-Tyrosine Kinases Biological Actins Endocytosis Protein Structure Tertiary DNA-Binding Proteins Settore BIO/18 - Genetica Mutagenesis Site-Directed Transcription Factors Plasmids Protein Binding Schizosaccharomyces pombe Proteins Peptides Mutagenesis Tertiary |
| Zdroj: | The Journal of biological chemistry. 277(7) |
| ISSN: | 0021-9258 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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