tion. In the latter, Itch WW domains interact with other proteins and can contribute to their ubiquitination and degradation. E3 ubiquitin ligases can also be phosphorylated at the WW sequence, thereby inhibiting their interactions. Our results suggest that while the WW domain is phosphorylated in a key residue for ligand recognition, such as T30, ligands are not efficiently recognised and targets can prevent their degradation. Thus, the initial change in conformation described in Itch after JNK1 activation could be the first step of a cascade of events that may include a WW domain phosphorylation/dephosphoylation equilibrium, thus providing a mechanism of target survival though inhibition of degradation. Further studies in vivo on whether phoshorylation can occur in these ancillary domains will make a crucial contribution to our understanding of activation and repression mechanisms of ubiquitin ligases. Structural work is underway to characterise the intramolecular interactions at an atomic level.
Set-up of the 600 MHz spectrometer
Publications
Research Networks and Grants
Brucet M, Querol-Audí J, Serra M, Ramirez-Espain X, Bertlik K, Ruiz L, Lloberas J, Macias MJ, Fita I and Celada A. Structure of the dimeric exonuclease TREX1 in complex with DNA displays a proline-rich binding site for WW Domains. J Biol Chem, 282(19), 14547-57 (2007)
Aplicación de la RMN a la determinación y caracterización dinámica de estructuras de proteínas, a la identificación de ligandos y a la caracterización de los correspondientes complejos Ministerio de Educación y Ciencia, GEN2003-20642-C09-04: 2004-2007 Research Director: Ernest Giralt
Macias MJ, Teijido O, Zifarelli G, Martin-Malpartida P, Ramirez-Espain X, Zorzano A, Palacín M, Pusch M and Estévez R. Myotonia-related mutations in the distal C-terminus of ClC-1 and ClC-0 chloride channels affect the structure of a poly-proline helix. Biochem J, 403(1), 79-87 (2007) Morales B, Ramirez-Espain X, Shaw AZ, Martin-Malpartida P, Yraola F, Sánchez-Tilló E, Farrera C, Celada A, Royo M and Macias MJ. NMR structural studies of the ItchWW3 domain reveal that phosphorylation at T30 inhibits the interaction with PPxY-containing ligands. Structure, 15(4), 473-83 (2007) Ramirez-Espain X, Ruiz L, Martin-Malpartida P, Oschkinat H and Macias MJ. Structural characterisation of a new binding motif and a novel binding mode in group 2 WW domains. J Mol Biol, 373(5), 1255-68 (2007)
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At the end of October, IRB Barcelona acquired a Bruker 600 MHz (14.1 Tesla) UltraShield magnet. Three weeks after its arrival, technical and scientific installations were finished. At present, we are running the standard set of triple resonance experiments to test the performance of this equipment. To date, the spectrometer is working as expected and giving high quality data sets.
Determinación de estructuras de dominios FF de proteínas y de sus interacciones mediante la aplicación de la resonancia magnética nuclear multidimensional en solución Ministerio de Educación y Ciencia, BFU2005-06276 IP: 20052008 Research Director: Maria J Macias
Collaborations
Synthesis of peptides and phosphorylated WW domains Miriam Royo, Barcelona Science Park (Barcelona Spain)
2007 S cie ntif ic Re po r t S t ruct ural and Co mput ational Biology Programme