title: Distinct ubiquitin binding modes exhibited by SH3 domains: molecular determinants and functional implications.
abstract: SH3 domains constitute a new type of ubiquitin-binding domains. We previously showed that the third SH3 domain (SH3-C) of CD2AP binds ubiquitin in an alternative orientation. We have determined the structure of the complex between first CD2AP SH3 domain and ubiquitin and performed a structural and mutational analysis to decipher the determinants of the SH3-C binding mode to ubiquitin. We found that the Phe-to-Tyr mutation in CD2AP and in the homologous CIN85 SH3-C domain does not abrogate ubiquitin binding, in contrast to previous hypothesis and our findings for the first two CD2AP SH3 domains. The similar alternative binding mode of the SH3-C domains of these related adaptor proteins is characterised by a higher affinity to C-terminal extended ubiquitin molecules. We conclude that CD2AP/CIN85 SH3-C domain interaction with ubiquitin constitutes a new ubiquitin-binding mode involved in a different cellular function and thus changes the previously established mechanism of EGF-dependent CD2AP/CIN85 mono-ubiquitination.
authors.affiliation: Departamento de Química Física e Instituto de Biotecnología, Facultad de Ciencias, Universidad de Granada, Granada, Spain ; Department of Biochemistry, University of Oxford, Oxford, United Kingdom.
authors.forename: Jose L
authors.initials: JL
authors.lastname: Ortega Roldan
authors.forename: Salvador
authors.initials: S
authors.lastname: Casares
authors.forename: Malene
authors.initials: M
authors.lastname: Ringkjøbing Jensen
authors.forename: Nayra
authors.initials: N
authors.lastname: Cárdenes
authors.forename: Jerónimo
authors.initials: J
authors.lastname: Bravo
authors.forename: Martin
authors.initials: M
authors.lastname: Blackledge
authors.forename: Ana I
authors.initials: AI
authors.lastname: Azuaga
authors.forename: Nico A J
authors.initials: NA
authors.lastname: van Nuland
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D009154:Mutation; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D025801:Ubiquitin / Q000737:chemistry* / Q000378:metabolism; D018909:src Homology Domains
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C114545:CD2-associated protein; D003598:Cytoskeletal Proteins; D025801:Ubiquitin
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