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Roles of the SH2 and SH3 domains in the regulation of neuronal Src kinase functions
Author(s) -
Groveman Bradley R.,
Xue Sheng,
Marin Vedrana,
Xu Jindong,
Ali Mohammad K.,
Bienkiewicz Ewa A.,
Yu XianMin
Publication year - 2011
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2010.07985.x
Subject(s) - sh2 domain , proto oncogene tyrosine protein kinase src , sh3 domain , tyrosine protein kinase csk , microbiology and biotechnology , tyrosine kinase , protein kinase domain , kinase , phosphorylation , biology , chemistry , biochemistry , signal transduction , gene , mutant
Previous studies demonstrated that intra‐domain interactions between Src family kinases (SFKs), stabilized by binding of the phosphorylated C‐terminus to the SH2 domain and/or binding of the SH2 kinase linker to the SH3 domain, lock the molecules in a closed conformation, disrupt the kinase active site, and inactivate SFKs. Here we report that the up‐regulation of N ‐methyl‐ d ‐aspartate receptors (NMDARs) induced by expression of constitutively active neuronal Src (n‐Src), in which the C‐terminus tyrosine is mutated to phenylalanine (n‐Src/Y535F), is significantly reduced by dysfunctions of the SH2 and/or SH3 domains of the protein. Furthermore, we found that dysfunctions of SH2 and/or SH3 domains reduce auto‐phosphorylation of the kinase activation loop, depress kinase activity, and decrease NMDAR phosphorylation. The SH2 domain plays a greater regulatory role than the SH3 domain. Our data also show that n‐Src binds directly to the C‐terminus of the NMDAR NR2A subunit in vitro , with a K D of 108.2 ± 13.3 n m . This binding is not Src kinase activity‐dependent, and dysfunctions of the SH2 and/or SH3 domains do not significantly affect the binding. These data indicate that the SH2 and SH3 domains may function to promote the catalytic activity of active n‐Src, which is important in the regulation of NMDAR functions. Structured digital abstract• MINT‐8074560 : NR2A (uniprotkb: Q00959 ) binds ( MI:0407 ) to n‐Src (uniprotkb: P05480 ) by surface plasmon resonance ( MI:0107 ) • MINT‐8074641 , MINT‐8074668 , MINT‐8074679 , MINT‐8074693 , MINT‐8074813 : n‐Src (uniprotkb: P05480 ) and n‐Src (uniprotkb: P05480 ) phosphorylate ( MI:0217 ) by protein kinase assay ( MI:0424 ) • MINT‐8074576 , MINT‐8074726 , MINT‐8074741 , MINT‐8074777 : n‐Src (uniprotkb: P05480 ) phosphorylates ( MI:0217 ) NR2A (uniprotkb: Q00959 ) by protein kinase assay ( MI:0424 )