title: Identification of phosphorylation sites within the SH3 domains of Tec family tyrosine kinases.
abstract: Tec family protein tyrosine kinases (TFKs) play a central role in hematopoietic cellular signaling. Initial activation takes place through specific tyrosine phosphorylation situated in the activation loop. Further activation occurs within the SH3 domain via a transphosphorylation mechanism, which for Bruton's tyrosine kinase (Btk) affects tyrosine 223. We found that TFKs phosphorylate preferentially their own SH3 domains, but differentially phosphorylate other member family SH3 domains, whereas non-related SH3 domains are not phosphorylated. We demonstrate that SH3 domains are good and reliable substrates. We observe that transphosphorylation is selective not only for SH3 domains, but also for dual SH3SH2 domains. However, the dual domain is phosphorylated more effectively. The major phosphorylation sites were identified as conserved tyrosines, for Itk Y180 and for Bmx Y215, both sites being homologous to the Y223 site in Btk. There is, however, one exception because the Tec-SH3 domain is phosphorylated at a non-homologous site, nevertheless a conserved tyrosine, Y206. Consistent with these findings, the 3D structures for SH3 domains point out that these phosphorylated tyrosines are located on the ligand-binding surface. Because a number of Tec family kinases are coexpressed in cells, it is possible that they could regulate the activity of each other through transphosphorylation.
authors.affiliation: Karolinska Institutet, Clinical Research Center (CRC) at Novum, Huddinge University Hospital, SE-141 86, Huddinge, Sweden. beston.nore@crc.ki.se
authors.forename: Beston F
authors.initials: BF
authors.lastname: Nore
authors.forename: Pekka T
authors.initials: PT
authors.lastname: Mattsson
authors.forename: Per
authors.initials: P
authors.lastname: Antonsson
authors.forename: Carl-Magnus
authors.initials: CM
authors.lastname: Bäckesjö
authors.forename: Anna
authors.initials: A
authors.lastname: Westlund
authors.forename: Johan
authors.initials: J
authors.lastname: Lennartsson
authors.forename: Henrik
authors.initials: H
authors.lastname: Hansson
authors.forename: Peter
authors.initials: P
authors.lastname: Löw
authors.forename: Lars
authors.initials: L
authors.lastname: Rönnstrand
authors.forename: C I Edvard
authors.initials: CI
authors.lastname: Smith
mesh_terms: D000077329:Agammaglobulinaemia Tyrosine Kinase; D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D003001:Cloning, Molecular; D006801:Humans; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis; D016415:Sequence Alignment; D018909:src Homology Domains / Q000235:genetics / Q000502:physiology*
chemical_list: D011994:Recombinant Proteins; C492367:BMX protein, human; C089848:Tec protein-tyrosine kinase; D011505:Protein-Tyrosine Kinases; D000077329:Agammaglobulinaemia Tyrosine Kinase; C000625949:BTK protein, human; C080813:emt protein-tyrosine kinase