TY - JOUR
T1 - Fyn phosphorylates human MAP-2c on tyrosine 67
AU - Zamora-Leon, S. Pilar
AU - Bresnick, Anne
AU - Backer, Jonathan M.
AU - Shafit-Zagardo, Bridget
PY - 2005/1/21
Y1 - 2005/1/21
N2 - The Src homology 3 (SH3)domain of Fyn binds to a conserved PXXP motif on microtubule-associated protein-2. Co-transfections into COS7 cells and in vitro kinase assays performed with Fyn and wild-type, or mutant MAP-2c, determined that Fyn phosphorylated MAP-2c on tyrosine 67. The phosphorylation generated a consensus sequence for the binding of the SH2 domain of Grb2 (pYSN). Pull-down assays with SH2-Grb2 from human fetal brain homogenates, and co-immunoprecipitation of Grb2 and MAP-2 confirmed the interaction in vivo, and demonstrated that MAP-2c is tyrosine-phosphorylateti in human fetal brain. Filter overlay assays confirmed that the SH2 domain of Grb2 binds to human MAP-2c following incubation with active Fyn. Enzyme-linked immunosorbent assays confirmed the interaction between the SH2 domain of Grb2 and a tyrosine-phosphorylated MAP-2 peptide spanning the PY67SN motif. Thus, MAP-2c can directly recruit multiple signaling proteins important for central nervous system development.
AB - The Src homology 3 (SH3)domain of Fyn binds to a conserved PXXP motif on microtubule-associated protein-2. Co-transfections into COS7 cells and in vitro kinase assays performed with Fyn and wild-type, or mutant MAP-2c, determined that Fyn phosphorylated MAP-2c on tyrosine 67. The phosphorylation generated a consensus sequence for the binding of the SH2 domain of Grb2 (pYSN). Pull-down assays with SH2-Grb2 from human fetal brain homogenates, and co-immunoprecipitation of Grb2 and MAP-2 confirmed the interaction in vivo, and demonstrated that MAP-2c is tyrosine-phosphorylateti in human fetal brain. Filter overlay assays confirmed that the SH2 domain of Grb2 binds to human MAP-2c following incubation with active Fyn. Enzyme-linked immunosorbent assays confirmed the interaction between the SH2 domain of Grb2 and a tyrosine-phosphorylated MAP-2 peptide spanning the PY67SN motif. Thus, MAP-2c can directly recruit multiple signaling proteins important for central nervous system development.
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U2 - 10.1074/jbc.M411380200
DO - 10.1074/jbc.M411380200
M3 - Article
C2 - 15536091
AN - SCOPUS:12544250513
SN - 0021-9258
VL - 280
SP - 1962
EP - 1970
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 3
ER -