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A novel peptide–SH3 interaction
Author(s) -
Mongioví Adriana Maria,
Romano Pascale R.,
Panni Simona,
Mendoza Manuel,
Wong William T.,
Musacchio Andrea,
Cesareni Gianni,
Paolo Di Fiore Pier
Publication year - 1999
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/18.19.5300
Subject(s) - sh3 domain , biology , subfamily , consensus sequence , binding site , peptide sequence , phage display , alanine , plasma protein binding , tyrosine , sequence alignment , peptide , peptide library , computational biology , biochemistry , receptor tyrosine kinase , amino acid , gene , kinase
SH3 domains constitute a family of protein–protein interaction modules that bind to peptides displaying an X‐proline‐X‐X‐proline (XPXXP) consensus. We report that the SH3 domain of Eps8, a substrate of receptor and non‐receptor tyrosine kinases, displays a novel and unique binding preference. By a combination of approaches including (i) screening of phage‐displayed random peptide libraries, (ii) mapping of the binding regions on three physiological interactors of Eps8, (iii) alanine scanning of binding peptides and (iv) in vitro cross‐linking, we demonstrate that a proline‐X‐X‐aspartate‐tyrosine (PXXDY) consensus is indispensable for binding to the SH3 domain of Eps8. Screening of the Expressed Sequence Tags database allowed the identification of three Eps8‐related genes, whose SH3s also display unusual binding preferences and constitute a phylogenetically distinct subfamily within the SH3 family. Thus, Eps8 identifies a novel family of SH3‐containing proteins that do not bind to canonical XPXXP‐containing peptides, and that establish distinct interactions in the signaling network.