Structure of the Eps15–stonin2 complex provides a molecular explanation for EH-domain ligand specificity
Open Access
- 17 January 2008
- journal article
- research article
- Published by Springer Nature in The EMBO Journal
- Vol. 27 (3), 558-569
- https://doi.org/10.1038/sj.emboj.7601980
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
This publication has 50 references indexed in Scilit:
- Functional Analysis of AP-2 α and μ2 SubunitsMolecular Biology of the Cell, 2006
- Role of the AP2 β-Appendage Hub in Recruiting Partners for Clathrin-Coated Vesicle AssemblyPLoS Biology, 2006
- Evolving nature of the AP2 α-appendage hub during clathrin-coated vesicle endocytosisThe EMBO Journal, 2004
- Dual Engagement Regulation of Protein Interactions with the AP-2 Adaptor α AppendageJournal of Biological Chemistry, 2004
- Stonin 2The Journal of cell biology, 2001
- Solution Structure of the Reps1 EH Domain and Characterization of Its Binding to NPF Target Sequences,Biochemistry, 2001
- AQUA and PROCHECK-NMR: Programs for checking the quality of protein structures solved by NMRJournal of Biomolecular NMR, 1996
- NMRPipe: A multidimensional spectral processing system based on UNIX pipesJournal of Biomolecular NMR, 1995
- SH3 Domains: Minding your p's and q'sCurrent Biology, 1995
- SPXX, a frequent sequence motif in gene regulatory proteinsJournal of Molecular Biology, 1989