Premium
Involvement of gangliosides in the process of Cbp/ PAG phosphorylation by L yn in developing cerebellar growth cones
Author(s) -
SekinoSuzuki Naoko,
Yuyama Kohei,
Miki Toshiaki,
Kaneda Mizuho,
Suzuki Hidenori,
Yamamoto Naomasa,
Yamamoto Tadashi,
Oneyama Chitose,
Okada Masato,
Kasahara Kohji
Publication year - 2013
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/jnc.12040
Subject(s) - lyn , immunoprecipitation , phosphorylation , tyrosine phosphorylation , ganglioside , immunoelectron microscopy , microbiology and biotechnology , biology , growth cone , cerebellum , western blot , biochemistry , proto oncogene tyrosine protein kinase src , chemistry , antibody , endocrinology , immunology , axon , gene
Abstract The association of gangliosides with specific proteins in the central nervous system was examined by coimmunoprecipitation with an anti‐ganglioside antibody. The monoclonal antibody to the ganglioside GD 3 (R24) immunoprecipitated the Csk (C‐terminal src kinase)‐binding protein (Cbp). Sucrose density gradient analysis showed that Cbp of rat cerebellum was detected in detergent‐resistant membrane ( DRM ) raft fractions. R24 treatment of the rat primary cerebellar cultures induced Lyn activation and tyrosine phosphorylation of Cbp. Treatment with anti‐ganglioside GD 1b antibody also induced tyrosine phosphorylation. Furthermore, over‐expressions of Lyn and Cbp in Chinese hamster ovary ( CHO ) cells resulted in tyrosine 314 phosphorylation of Cbp, which indicates that Cbp is a substrate for Lyn. Immunoblotting analysis showed that the active form of Lyn and the Tyr314‐phosphorylated form of Cbp were highly accumulated in the DRM raft fraction prepared from the developing cerebellum compared with the DRM raft fraction of the adult one. In addition, Lyn and the Tyr314‐phosphorylated Cbp were highly concentrated in the growth cone fraction prepared from the developing cerebellum. Immunoelectron microscopy showed that Cbp and GAP ‐43, a growth cone marker, are localized in the same vesicles of the growth cone fraction. These results suggest that Cbp functionally associates with gangliosides on growth cone rafts in developing cerebella.