title: Complex formation of EphB1/Nck/Caskin1 leads to tyrosine phosphorylation and structural changes of the Caskin1 SH3 domain.
abstract: BACKGROUND
  Scaffold proteins have an important role in the regulation of signal propagation. These proteins do not possess any enzymatic activity but can contribute to the formation of multiprotein complexes. Although scaffold proteins are present in all cell types, the nervous system contains them in the largest amount. Caskin proteins are typically present in neuronal cells, particularly, in the synapses. However, the signaling mechanisms by which Caskin proteins are regulated are largely unknown.
  
  
  RESULTS
  Here we demonstrate that EphB1 receptor tyrosine kinase can recruit Caskin1 through the adaptor protein Nck. Upon activation of the receptor kinase, the SH2 domain of Nck binds to one of its tyrosine residues, while Nck SH3 domains interact with the proline-rich domain of Caskin1. Complex formation of the receptor, adaptor and scaffold proteins results in the tyrosine phosphorylation of Caskin1 on its SH3 domain. The phosphorylation sites were identified by mass-spectrometry as tyrosines 296 and 336. To reveal the structural consequence of this phosphorylation, CD spectroscopy was performed. This measurement suggests that upon tyrosine phosphorylation the structure of the Caskin1 SH3 domain changes significantly.
  
  
  CONCLUSIONS
  Taken together, we propose that the scaffold protein Caskin1 can form a complex with the EphB1 tyrosine kinase via the Nck protein as a linker. Complex formation results in tyrosine phosphorylation of the Caskin1 SH3 domain. Although we were not able to identify any physiological partner of the SH3 domain so far, we could demonstrate that phosphorylation on conserved tyrosine residues results in marked changes in the structure of the SH3 domain.
authors.affiliation: From the Department of Medical Chemistry, Semmelweis University Medical School, Budapest, 1094, Hungary. buday.laszlo@ttk.mta.hu.
authors.forename: Szabolcs
authors.initials: S
authors.lastname: Pesti
authors.forename: Annamária
authors.initials: A
authors.lastname: Balázs
authors.forename: Roopesh
authors.initials: R
authors.lastname: Udupa
authors.forename: Beáta
authors.initials: B
authors.lastname: Szabó
authors.forename: Anna
authors.initials: A
authors.lastname: Fekete
authors.forename: Gábor
authors.initials: G
authors.lastname: Bőgel
authors.forename: László
authors.initials: L
authors.lastname: Buday