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Association of gangliosides to fibroblasts in culture: A study performed with GM1 [14C]-labelled at the sialic acid acetyl group

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Abstract

The preparation of a GM1-ganglioside (GM1) [14C]-labelled in the sialic acid residue is reported. This can be obtained by re-N-acetylation in the presence of [1-14C]-acetic anhydride, of a GM1 derivative de-N-acetylated specifically on the sialic acid residue by alkaline hydrolysis of GM1 with tetramethylammonium hydroxide. The radiolabelled GM1 is utilized to investigate the binding properties and the mode of interaction of GM1 with cultured fibroblasts. Three different forms of association (one “serum-removable”, one “trypsin-removable” and one “trypsin-stable”) have been recognized to occur in a way that depended on cell culture conditions (presence or absence of fetal calf serum), ganglioside concentration (from, 5×10−9 M to 10−4 M) and incubation time (up to 24 h). Some metabolic modifications of GM1 during the period of high cell viability were also investigated.

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Abbreviations

GM1:

GM1-ganglioside, II3NeuAc-GgOse4Cer

FCS:

fetal calf serum

EMEM:

Eaglés Minimum Essential Medium with Earlés salts

PBS:

Dulbecco phosphate buffered saline without calcium and magnesium

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Authors and Affiliations

  1. Study Center for the Functional Biochemistry of Brain Lipids, Department of Biological Chemistry, The Medical School, University of Milan, Via Saldini 50, 20133, Milan, Italy

    Vanna Chigorno, Marina Pitto, Gloria Cardace, Domenico Acquotti, Sandro Sonnino, Riccardo Ghidoni & Guido Tettamanti

  2. Department of Chemistry, Fidia Research Laboratories, Abano Terme, Italy

    Günther Kirschner

Authors
  1. Vanna Chigorno
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  2. Marina Pitto
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  3. Gloria Cardace
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  4. Domenico Acquotti
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  5. Günther Kirschner
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  6. Sandro Sonnino
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  7. Riccardo Ghidoni
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  8. Guido Tettamanti
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Chigorno, V., Pitto, M., Cardace, G. et al. Association of gangliosides to fibroblasts in culture: A study performed with GM1 [14C]-labelled at the sialic acid acetyl group. Glycoconjugate J 2, 279–291 (1985). https://doi.org/10.1007/BF01049274

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  • DOI: https://doi.org/10.1007/BF01049274

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