Identification of PSD-93 as a substrate for the Src family tyrosine kinase Fyn

Shigeyuki Nada, Takaki Shima, Hiroyuki Yanai, Holger Husi, Seth G.N. Grant, Masato Okada, Tetsu Akiyama

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39 Citations (Scopus)

Abstract

In order to study the role of tyrosine kinase signaling in the post-synaptic density (PSD), tyrosine-phosphorylated proteins associated with the PSD-95/NMDA receptor complex were analyzed. The NMDA receptor complex from the mouse brain was successfully solubilized with deoxycholate and immunopurified with anti-PSD-95 or anti-phosphotyrosine antibody. Immunoblot analyses revealed that the predominantly tyrosine-phosphorylated proteins in the NMDA receptor complex are the NR2A/B subunits and a novel 120 kDa protein. Purification and microsequencing analysis showed that the 120 kDa protein is mouse PSD-93/Chapsyn-110. Recombinant PSD-93 was phosphorylated by Fyn in vitro, and Tyr-384 was identified as a major phosphorylation site. Tyrosine phosphorylation of PSD-93 was greatly reduced in brain tissue from Fyn-deficient mice compared with wild-type mice. Furthermore, an N-terminal palmitoylation signal of PSD-93 was found to be essential for its anchoring to the membrane, where Fyn is also localized. In COS7 cells, exogenously expressed PSD-93 was phosphorylated, dependent on its membrane localization. In addition, tyrosine-phosphorylated PSD-93 was able to bind to Csk, a negative regulator of Src family kinases, in vitro as well as in a brain lysate. These results suggest that PSD-93 serves as a membrane-anchored substrate of Fyn and plays a role in the regulation of Fyn-mediated modification of NMDA receptor function.
Original languageEnglish
Pages (from-to)47610-47621
Number of pages12
JournalThe Journal of Biological Chemistry
Volume278
Issue number48
DOIs
Publication statusPublished - 1 Sep 2003

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    Nada, S., Shima, T., Yanai, H., Husi, H., Grant, S. G. N., Okada, M., & Akiyama, T. (2003). Identification of PSD-93 as a substrate for the Src family tyrosine kinase Fyn. The Journal of Biological Chemistry, 278(48), 47610-47621. https://doi.org/10.1074/jbc.M303873200