The first two authors contributed equally to the experimental work.
Specific interactions of neuronal focal adhesion kinase isoforms with Src kinases and amphiphysin
Article first published online: 9 JAN 2003
Journal of Neurochemistry
Volume 84, Issue 2, pages 253–265, January 2003
How to Cite
Messina, S., Onofri, F., Bongiorno-Borbone, L., Giovedì, S., Valtorta, F., Girault, J.-A. and Benfenati, F. (2003), Specific interactions of neuronal focal adhesion kinase isoforms with Src kinases and amphiphysin. Journal of Neurochemistry, 84: 253–265. doi: 10.1046/j.1471-4159.2003.01519.x
- Issue published online: 9 JAN 2003
- Article first published online: 9 JAN 2003
- Received June 19, 2002; revised manuscript received September 27, 2002; accepted October 6, 2002.
- nerve terminals;
- SH2 domains;
- SH3 domains;
- tyrosine phosphorylation
Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that activates Src family kinases via SH2- and SH3-mediated interactions. Specific FAK isoforms (FAK+), responsive to depolarization and neurotransmitters, are enriched in neurons. We analyzed the interactions of endogenous FAK+ and recombinant FAK+ isoforms containing amino acid insertions (boxes 6,7,28) with an array of SH3 domains and the c-Src SH2/SH3 domain tandem. Endogenous FAK+ bound specifically to the SH3 domains of c-Src (but not n-Src), Fyn, Yes, phosphtidylinositol-3 kinase, amphiphysin II, amphiphysin I, phospholipase Cγ and NH2-terminal Grb2. The inclusion of boxes 6,7 was associated with a significant decrease in the binding of FAK+ to the c-Src and Fyn SH3 domains, and a significant increase in the binding to the Src SH2 domain, as a consequence of the higher phosphorylation of Tyr-397. The novel interaction with the amphiphysin SH3 domain, involving the COOH-terminal proline-rich region of FAK, was confirmed by coimmunoprecipitation of the two proteins and a closely similar response to stimuli affecting the actin cytoskeleton. Moreover, an impairment of endocytosis was observed in synaptosomes after internalization of a proline-rich peptide corresponding to the site of interaction. The data account for the different subcellular distribution of FAK and Src kinases and the specific regulation of the transduction pathways linked to FAK activation in the brain and implicate FAK in the regulation of membrane trafficking in nerve terminals.