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

Research output: Contribution to journalArticle

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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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Microfilament Proteins
Functional analysis
Fungal Proteins
src Homology Domains
Structural analysis
Yeast
Actins
Ligands
Mutagenesis
Peptides
Bacteriophages
Phosphotransferases
Genes
Display devices
Throughput
Site-Directed Mutagenesis
Proteins
Actin Cytoskeleton
Saccharomyces cerevisiae
Genome

ASJC Scopus subject areas

  • Biochemistry

Cite this

Fazi, B., Cope, M. J. T. V., Douangamath, A., Ferracuti, S., Schirwitz, K., Zucconi, A., ... 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

Unusual binding properties of the SH3 domain of the yeast actin-binding protein Abp1. Structural and functional analysis. / Fazi, Barbara; Cope, M. Jamie T V; Douangamath, Alice; Ferracuti, Silvia; Schirwitz, Katja; Zucconi, Adriana; Drubin, David G.; Wilmanns, Matthias; Cesareni, Gianni; Castagnoli, Luisa.

In: Journal of Biological Chemistry, Vol. 277, No. 7, 15.02.2002, p. 5290-5298.

Research output: Contribution to journalArticle

Fazi, B, Cope, MJTV, Douangamath, A, Ferracuti, S, Schirwitz, K, Zucconi, A, Drubin, DG, 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, vol. 277, no. 7, pp. 5290-5298. https://doi.org/10.1074/jbc.M109848200
Fazi, Barbara ; Cope, M. Jamie T V ; Douangamath, Alice ; Ferracuti, Silvia ; Schirwitz, Katja ; Zucconi, Adriana ; Drubin, David G. ; Wilmanns, Matthias ; Cesareni, Gianni ; Castagnoli, Luisa. / Unusual binding properties of the SH3 domain of the yeast actin-binding protein Abp1. Structural and functional analysis. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 7. pp. 5290-5298.
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