Article
- The EMBO Journal (2008) 27, 558 - 569
- doi:10.1038/sj.emboj.7601980
Published online: 17 January 2008
Subject Categories:
Structure of the Eps15–stonin2 complex provides a molecular explanation for EH-domain ligand specificity
Julia Rumpf1, Bernd Simon2, Nadja Jung3, Tanja Maritzen3, Volker Haucke3, Michael Sattler2,4,5 and Yvonne Groemping1
- Department of Biomolecular Mechanisms, Max-Planck-Institute for Medical Research, Heidelberg, Germany
- European Molecular Biology Laboratory, Heidelberg, Germany
- Institut für Chemie und Biochemie, Freie Universitaet Berlin, Berlin, Germany
- GSF-National Research Center for Environment and Health, Neuherberg, Germany
- Munich Center for Integrated Protein Science, Department Chemie, Technische Universität München, Garching, Germany
Correspondence to:
Yvonne Groemping, Department of Biomolecular Mechanisms, Max-Planck-Institute for Medical Research, Jahnstrasse 29, Heidelberg 69120, Germany. Tel.: +49 486 536; Fax: +49 486 585; E-mail: yvonne.groemping@mpimf-heidelberg.mpg.de
Received 2 August 2007; Accepted 17 December 2007
Abstract
Eps15 homology (EH) domain-containing proteins play a key regulatory role in intracellular membrane trafficking and cell signalling. EH domains serve as interaction platforms for short peptide motifs comprising the residues NPF within natively unstructured regions of accessory proteins. The EH–NPF interactions described thus far are of very low affinity and specificity. Here, we identify the presynaptic endocytic sorting adaptor stonin2 as a high-affinity ligand for the second EH domain (EH2) of the clathrin accessory protein Eps15. Calorimetric data indicate that both NPF motifs within stonin2 interact with EH2 simultaneously and with sub-micromolar affinity. The solution structure of this complex reveals that the first NPF motif binds to the conserved site on the EH domain, whereas the second motif inserts into a novel hydrophobic pocket. Our data show how combination of two EH-attachment sites provides a means for modulating specificity and allows discrimination from a large pool of potential binding partners containing NPF motifs.
Keywords:
- endocytosis,
- Eps15,
- protein–protein interactions,
- specificity,
- stonin2
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