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Fyn phosphorylates human MAP-2c on tyrosine 67
S Pilar Zamora-Leon1, Anne Bresnick, Jonathan M Backer
1Department of Pathology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
The Journal of Biological Chemistry
|November 13, 2004
Summary
Fyn kinase phosphorylates microtubule-associated protein-2 (MAP-2c), enabling Grb2 binding. This interaction recruits signaling proteins crucial for central nervous system development.
Area of Science:
- Molecular Biology
- Neuroscience
- Cell Signaling
Background:
- Microtubule-associated protein-2 (MAP-2) plays a role in neuronal development.
- Fyn kinase is involved in various signaling pathways.
- Grb2 acts as an adaptor protein in signal transduction.
Purpose of the Study:
- To investigate the interaction between Fyn kinase and MAP-2c.
- To determine if Fyn phosphorylation of MAP-2c affects Grb2 binding.
- To elucidate the role of this interaction in central nervous system development.
Main Methods:
- Co-transfection assays in COS7 cells.
- In vitro kinase assays using Fyn and MAP-2c variants.
- Pull-down assays with SH2-Grb2 and human fetal brain homogenates.
- Co-immunoprecipitation of Grb2 and MAP-2.
- Filter overlay assays.
- Enzyme-linked immunosorbent assays (ELISAs).
Main Results:
- Fyn kinase phosphorylates MAP-2c on tyrosine 67.
- Phosphorylation creates a binding site (pYSN) for the SH2 domain of Grb2.
- MAP-2c is tyrosine-phosphorylated in human fetal brain.
- The SH2 domain of Grb2 directly binds to phosphorylated MAP-2c.
- MAP-2c directly recruits signaling proteins via Grb2.
Conclusions:
- Fyn-mediated phosphorylation of MAP-2c is a key regulatory event.
- This phosphorylation facilitates the recruitment of Grb2, linking MAP-2c to downstream signaling pathways.
- MAP-2c serves as a scaffold for signaling proteins essential for CNS development.