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EMBO J. Nov 16, 1998; 17(22): 6541–6550.
PMCID: PMC1171001

Recognition specificity of individual EH domains of mammals and yeast.


The Eps homology (EH) domain is a recently described protein binding module that is found, in multiple or single copies, in several proteins in species as diverse as human and yeast. In this work, we have investigated the molecular details of recognition specificity mediated by this domain family by characterizing the peptide-binding preference of 11 different EH domains from mammal and yeast proteins. Ten of the eleven EH domains could bind at least some peptides containing an Asn-Pro-Phe (NPF) motif. By contrast, the first EH domain of End3p preferentially binds peptides containing an His-Thr/Ser-Phe (HT/SF) motif. Domains that have a low affinity for the majority of NPF peptides reveal some affinity for a third class of peptides that contains two consecutive amino acids with aromatic side chains (FW or WW). This is the case for the third EH domain of Eps15 and for the two N-terminal domains of YBL47c. The consensus sequences derived from the peptides selected from phage-displayed peptide libraries allows for grouping of EH domains into families that are characterized by different NPF-context preference. Finally, comparison of the primary sequence of EH domains with similar or divergent specificity identifies a residue at position +3 following a conserved tryptophan, whose chemical characteristics modulate binding preference.

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Selected References

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  • Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 1997 Sep 1;25(17):3389–3402. [PMC free article] [PubMed]
  • Bénédetti H, Raths S, Crausaz F, Riezman H. The END3 gene encodes a protein that is required for the internalization step of endocytosis and for actin cytoskeleton organization in yeast. Mol Biol Cell. 1994 Sep;5(9):1023–1037. [PMC free article] [PubMed]
  • Benmerah A, Bégue B, Dautry-Varsat A, Cerf-Bensussan N. The ear of alpha-adaptin interacts with the COOH-terminal domain of the Eps 15 protein. J Biol Chem. 1996 May 17;271(20):12111–12116. [PubMed]
  • Benmerah A, Lamaze C, Bègue B, Schmid SL, Dautry-Varsat A, Cerf-Bensussan N. AP-2/Eps15 interaction is required for receptor-mediated endocytosis. J Cell Biol. 1998 Mar 9;140(5):1055–1062. [PMC free article] [PubMed]
  • Carbone R, Fré S, Iannolo G, Belleudi F, Mancini P, Pelicci PG, Torrisi MR, Di Fiore PP. eps15 and eps15R are essential components of the endocytic pathway. Cancer Res. 1997 Dec 15;57(24):5498–5504. [PubMed]
  • Coda L, Salcini AE, Confalonieri S, Pelicci G, Sorkina T, Sorkin A, Pelicci PG, Di Fiore PP. Eps15R is a tyrosine kinase substrate with characteristics of a docking protein possibly involved in coated pits-mediated internalization. J Biol Chem. 1998 Jan 30;273(5):3003–3012. [PubMed]
  • de Beer T, Carter RE, Lobel-Rice KE, Sorkin A, Overduin M. Structure and Asn-Pro-Phe binding pocket of the Eps15 homology domain. Science. 1998 Aug 28;281(5381):1357–1360. [PubMed]
  • Dente L, Cesareni G, Cortese R. pEMBL: a new family of single stranded plasmids. Nucleic Acids Res. 1983 Mar 25;11(6):1645–1655. [PMC free article] [PubMed]
  • Dente L, Vetriani C, Zucconi A, Pelicci G, Lanfrancone L, Pelicci PG, Cesareni G. Modified phage peptide libraries as a tool to study specificity of phosphorylation and recognition of tyrosine containing peptides. J Mol Biol. 1997 Jun 27;269(5):694–703. [PubMed]
  • Di Fiore PP, Pelicci PG, Sorkin A. EH: a novel protein-protein interaction domain potentially involved in intracellular sorting. Trends Biochem Sci. 1997 Nov;22(11):411–413. [PubMed]
  • Fazioli F, Minichiello L, Matoskova B, Wong WT, Di Fiore PP. eps15, a novel tyrosine kinase substrate, exhibits transforming activity. Mol Cell Biol. 1993 Sep;13(9):5814–5828. [PMC free article] [PubMed]
  • Felici F, Castagnoli L, Musacchio A, Jappelli R, Cesareni G. Selection of antibody ligands from a large library of oligopeptides expressed on a multivalent exposition vector. J Mol Biol. 1991 Nov 20;222(2):301–310. [PubMed]
  • Haffner C, Takei K, Chen H, Ringstad N, Hudson A, Butler MH, Salcini AE, Di Fiore PP, De Camilli P. Synaptojanin 1: localization on coated endocytic intermediates in nerve terminals and interaction of its 170 kDa isoform with Eps15. FEBS Lett. 1997 Dec 15;419(2-3):175–180. [PubMed]
  • Iannolo G, Salcini AE, Gaidarov I, Goodman OB, Jr, Baulida J, Carpenter G, Pelicci PG, Di Fiore PP, Keen JH. Mapping of the molecular determinants involved in the interaction between eps15 and AP-2. Cancer Res. 1997 Jan 15;57(2):240–245. [PubMed]
  • Ikura M, Clore GM, Gronenborn AM, Zhu G, Klee CB, Bax A. Solution structure of a calmodulin-target peptide complex by multidimensional NMR. Science. 1992 May 1;256(5057):632–638. [PubMed]
  • Meador WE, Means AR, Quiocho FA. Target enzyme recognition by calmodulin: 2.4 A structure of a calmodulin-peptide complex. Science. 1992 Aug 28;257(5074):1251–1255. [PubMed]
  • Munn AL, Stevenson BJ, Geli MI, Riezman H. end5, end6, and end7: mutations that cause actin delocalization and block the internalization step of endocytosis in Saccharomyces cerevisiae. Mol Biol Cell. 1995 Dec;6(12):1721–1742. [PMC free article] [PubMed]
  • Pawson T, Scott JD. Signaling through scaffold, anchoring, and adaptor proteins. Science. 1997 Dec 19;278(5346):2075–2080. [PubMed]
  • Rickles RJ, Botfield MC, Weng Z, Taylor JA, Green OM, Brugge JS, Zoller MJ. Identification of Src, Fyn, Lyn, PI3K and Abl SH3 domain ligands using phage display libraries. EMBO J. 1994 Dec 1;13(23):5598–5604. [PMC free article] [PubMed]
  • Roos J, Kelly RB. Dap160, a neural-specific Eps15 homology and multiple SH3 domain-containing protein that interacts with Drosophila dynamin. J Biol Chem. 1998 Jul 24;273(30):19108–19119. [PubMed]
  • Salcini AE, Confalonieri S, Doria M, Santolini E, Tassi E, Minenkova O, Cesareni G, Pelicci PG, Di Fiore PP. Binding specificity and in vivo targets of the EH domain, a novel protein-protein interaction module. Genes Dev. 1997 Sep 1;11(17):2239–2249. [PMC free article] [PubMed]
  • Songyang Z, Shoelson SE, Chaudhuri M, Gish G, Pawson T, Haser WG, King F, Roberts T, Ratnofsky S, Lechleider RJ, et al. SH2 domains recognize specific phosphopeptide sequences. Cell. 1993 Mar 12;72(5):767–778. [PubMed]
  • Songyang Z, Shoelson SE, McGlade J, Olivier P, Pawson T, Bustelo XR, Barbacid M, Sabe H, Hanafusa H, Yi T, et al. Specific motifs recognized by the SH2 domains of Csk, 3BP2, fps/fes, GRB-2, HCP, SHC, Syk, and Vav. Mol Cell Biol. 1994 Apr;14(4):2777–2785. [PMC free article] [PubMed]
  • Songyang Z, Fanning AS, Fu C, Xu J, Marfatia SM, Chishti AH, Crompton A, Chan AC, Anderson JM, Cantley LC. Recognition of unique carboxyl-terminal motifs by distinct PDZ domains. Science. 1997 Jan 3;275(5296):73–77. [PubMed]
  • Sparks AB, Quilliam LA, Thorn JM, Der CJ, Kay BK. Identification and characterization of Src SH3 ligands from phage-displayed random peptide libraries. J Biol Chem. 1994 Sep 30;269(39):23853–23856. [PubMed]
  • Tan PK, Howard JP, Payne GS. The sequence NPFXD defines a new class of endocytosis signal in Saccharomyces cerevisiae. J Cell Biol. 1996 Dec;135(6 Pt 2):1789–1800. [PMC free article] [PubMed]
  • Tang HY, Cai M. The EH-domain-containing protein Pan1 is required for normal organization of the actin cytoskeleton in Saccharomyces cerevisiae. Mol Cell Biol. 1996 Sep;16(9):4897–4914. [PMC free article] [PubMed]
  • Tang HY, Munn A, Cai M. EH domain proteins Pan1p and End3p are components of a complex that plays a dual role in organization of the cortical actin cytoskeleton and endocytosis in Saccharomyces cerevisiae. Mol Cell Biol. 1997 Aug;17(8):4294–4304. [PMC free article] [PubMed]
  • Tebar F, Sorkina T, Sorkin A, Ericsson M, Kirchhausen T. Eps15 is a component of clathrin-coated pits and vesicles and is located at the rim of coated pits. J Biol Chem. 1996 Nov 15;271(46):28727–28730. [PubMed]
  • van Delft S, Schumacher C, Hage W, Verkleij AJ, van Bergen en Henegouwen PM. Association and colocalization of Eps15 with adaptor protein-2 and clathrin. J Cell Biol. 1997 Feb 24;136(4):811–821. [PMC free article] [PubMed]
  • Wendland B, Emr SD. Pan1p, yeast eps15, functions as a multivalent adaptor that coordinates protein-protein interactions essential for endocytosis. J Cell Biol. 1998 Apr 6;141(1):71–84. [PMC free article] [PubMed]
  • Wong WT, Schumacher C, Salcini AE, Romano A, Castagnino P, Pelicci PG, Di Fiore PP. A protein-binding domain, EH, identified in the receptor tyrosine kinase substrate Eps15 and conserved in evolution. Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9530–9534. [PMC free article] [PubMed]
  • Yamaguchi A, Urano T, Goi T, Feig LA. An Eps homology (EH) domain protein that binds to the Ral-GTPase target, RalBP1. J Biol Chem. 1997 Dec 12;272(50):31230–31234. [PubMed]

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