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

Barbara Fazi, M. Jamie T V Cope, Alice Douangamath, Silvia Ferracuti, Katja Schirwitz, Adriana Zucconi, David G. Drubin, Matthias Wilmanns, Gianni Cesareni, Luisa Castagnoli

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Abstract

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-Å 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.

Original languageEnglish
Pages (from-to)5290-5298
Number of pages9
JournalJournal of Biological Chemistry
Volume277
Issue number7
DOIs
Publication statusPublished - Feb 15 2002

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ASJC Scopus subject areas

  • Biochemistry

Cite this

Fazi, B., Cope, M. J. T. V., Douangamath, A., Ferracuti, S., Schirwitz, K., Zucconi, A., Drubin, D. G., Wilmanns, M., Cesareni, G., & Castagnoli, L. (2002). Unusual binding properties of the SH3 domain of the yeast actin-binding protein Abp1. Structural and functional analysis. Journal of Biological Chemistry, 277(7), 5290-5298. https://doi.org/10.1074/jbc.M109848200