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An anti-sulfatide antibody O4 immunoprecipitates sulfatide rafts including Fyn, Lyn and the G protein α subunit in rat primary immature oligodendrocytes

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Abstract

The association of sulfatide with specific proteins in oligodendrocytes was examined by co-immunoprecipitation with an anti-sulfatide antibody. Protein kinase activity was detected in precipitates with a monoclonal antibody to sulfatide (O4) from the rat primary immature oligodendrocytes. We conducted in vitro kinase assay of tyrosine phosphorylated proteins of 80, 59, 56, 53 and 40 kDa by gel electrophoresis. Of these proteins, the proteins of 59 kDa and 53/56 kDa were identified as the Src family tyrosine kinases Fyn and Lyn on the basis of their sequential immunoprecipitation with anti-Fyn and anti-Lyn antibodies, respectively. The 40 kDa protein was identified as the α subunit of the heterotrimeric G protein. These observations suggest that O4 immunoprecipitates sulfatide rafts including Fyn, Lyn and the α subunit of the heterotrimeric G protein.

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Acknowledgments

This work was supported by JSPS KAKENHI, Grant Number 23570182, for Scientific Research (C).

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The authors declare no conflicts of interest.

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Authors and Affiliations

  1. Laboratory of Biomembrane, Tokyo Metropolitan Institute of Medical Science, Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan

    Toshiaki Miki, Mizuho Kaneda, Kazuko Iida, Go Hasegawa & Kohji Kasahara

  2. Lipid Metabolism Project, Tokyo Metropolitan Institute of Medical Science, Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan

    Makoto Murakami

  3. Faculty of Pharmaceutical Sciences, Ohu University, Koriyama, Fukushima, 963-8611, Japan

    Naomasa Yamamoto

  4. Center for Kampo Medicine, Keio University School of Medicine, Shinjyuku-ku, Tokyo, 160-8582, Japan

    Hiroaki Asou

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  1. Toshiaki Miki
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Correspondence to Kohji Kasahara.

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Miki, T., Kaneda, M., Iida, K. et al. An anti-sulfatide antibody O4 immunoprecipitates sulfatide rafts including Fyn, Lyn and the G protein α subunit in rat primary immature oligodendrocytes. Glycoconj J 30, 819–823 (2013). https://doi.org/10.1007/s10719-013-9487-5

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