Carregant...
Fitxers
Tipus de document
ArticleVersió
Versió acceptadaData de publicació
Tots els drets reservats
Si us plau utilitzeu sempre aquest identificador per citar o enllaçar aquest document: https://hdl.handle.net/2445/138577
Multi-phosphorylation of the intrinsically disordered unique domain of c-Src studied by in-cell and real-time NMR
Títol de la revista
Director/Tutor
ISSN de la revista
Títol del volum
Recurs relacionat
Resum
Intrinsically disordered regions (IDRs) are preferred sites for post-translational modifications essential for regulating protein function. The enhanced local mobility of IDRs facilitates their observation by NMR spectroscopy in vivo. Phosphorylation events can occur at multiple sites and respond dynamically to changes in kinase-phosphatase networks. Here we used real-time NMR spectroscopy to study the effect of kinases and phosphatases present in Xenopus oocytes and egg extracts on the phosphorylation state of the 'unique domain' of c-Src. We followed the phosphorylation of S17 in oocytes, and of S17, S69, and S75 in egg extracts by NMR spectroscopy, MS, and western blotting. Addition of specific kinase inhibitors showed that S75 and S69 are phosphorylated by CDKs (cyclin-dependent kinases) differently from Cdk1. Moreover, although PKA (cAMP-dependent protein kinase) can phosphorylate S17 in vitro, this was not the major S17 kinase in egg extracts. Changes in PKA activity affected the phosphorylation levels of CDK-dependent sites, thus suggesting indirect effects of kinase-phosphatase networks. This study provides a proof-of-concept of the use of real-time in vivo NMR spectroscopy to characterize kinase/phosphatase effects on intrinsically disordered regulatory domains.
Matèries (anglès)
Citació
Citació
AMATA, Irene, MAFFEI, Mariano, IGEA, Ana, GAY I MARÍN, Marina, VILASECA CASAS, Marta, NEBREDA, Àngel r., PONS VALLÈS, Miquel. Multi-phosphorylation of the intrinsically disordered unique domain of c-Src studied by in-cell and real-time NMR. _ChemBioChem_. 2013. Vol. 14, núm. 14, pàgs. 1820-1827. [consulta: 20 de gener de 2026]. ISSN: 1439-4227. [Disponible a: https://hdl.handle.net/2445/138577]