J. Biochem, 1988, Vol. 104, No. 4 498-503
© 1988 Japanese Biochemical Society
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A Novel, Carbohydrate Signal-Mediated Cell Surface Protein Phosphorylation: Ganglioside GQ1b Stimulates Ecto-Protein Kinase Activity on the Cell Surface of a Human Neuroblastoma Cell Line, GOTO2
*Department of Biochemistry, Faculty of Medicine,The University of Tokyo Bunkyo-ku, Tokyo 113
**Department of Neurobiology, Brain Research Institute, Niigata University Niigata, Niigata 951
2To whom correspondence should be addressed
A ganglioside-stimulated ecto.type protein phosphorylation system (ecto-Gg-kinase) was detected on the cell surface of a human neuroblastoma cell line (GOTO). When intact cells were incubated with [
-32]ATP at least 28 cell surface proteins were phosphorylated, as evident on SDS-PAGE (4-20%) analysis. Exogenously added gangliosides specifically stimulated the phosphorylation of at least three cell surface associated proteins of Mr = 64,000, 60,000, and 54,000. Phosphorylation was directed toward Thr and Ser residues, respectively, as revealed on acid hydrolysis followed by electrophoresis. GQ1b, at 5 nM, was the most potent among the several gangliosides tested and was more effective when added to cells before [-32]ATP administration. The simultaneous addition of an excess amount of the saccharide portion of GQ1b (oligo-GQ1b) inhibited the GQ1b-stimulated phosphorylation,indicating the necessity of the sialosaccharide moiety. These results strongly suggest that phosphorylation of the three proteins may be closely associated with the highly specific neuritogenic effect of GQ1b previously reported.
1 This work was supported in part by a Grant-in-Aid for the Promotion of Science and a Grant-in-Aid for Cancer Research, both from the Ministry of Education, Science and Culture of Japan. This work was also supported in part by Haraguchi Memorial Cancer Research Fund.
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