Fyn phosphorylates human MAP-2c on tyrosine 67

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1962-1970
Number of pages9
JournalJournal of Biological Chemistry
Volume280
Issue number3
DOIs
StatePublished - Jan 21 2005

Fingerprint

src Homology Domains
Tyrosine
Assays
Brain
Immunosorbents
Phosphorylation
Microtubule-Associated Proteins
Consensus Sequence
Neurology
Immunoprecipitation
Transfection
Phosphotransferases
Central Nervous System
Enzyme-Linked Immunosorbent Assay
Peptides
Enzymes
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Fyn phosphorylates human MAP-2c on tyrosine 67. / Zamora-Leon, S. Pilar; Bresnick, Anne R.; Backer, Jonathan M.; Shafit-Zagardo, Bridget.

In: Journal of Biological Chemistry, Vol. 280, No. 3, 21.01.2005, p. 1962-1970.

Research output: Contribution to journalArticle

@article{1a0cc0dc91304f52ada1d4e589b83171,
title = "Fyn phosphorylates human MAP-2c on tyrosine 67",
abstract = "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.",
author = "Zamora-Leon, {S. Pilar} and Bresnick, {Anne R.} and Backer, {Jonathan M.} and Bridget Shafit-Zagardo",
year = "2005",
month = "1",
day = "21",
doi = "10.1074/jbc.M411380200",
language = "English (US)",
volume = "280",
pages = "1962--1970",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "3",

}

TY - JOUR

T1 - Fyn phosphorylates human MAP-2c on tyrosine 67

AU - Zamora-Leon, S. Pilar

AU - Bresnick, Anne R.

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.

UR - http://www.scopus.com/inward/record.url?scp=12544250513&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=12544250513&partnerID=8YFLogxK

U2 - 10.1074/jbc.M411380200

DO - 10.1074/jbc.M411380200

M3 - Article

C2 - 15536091

AN - SCOPUS:12544250513

VL - 280

SP - 1962

EP - 1970

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 3

ER -