Association of gangliosides to fibroblasts in culture: A study performed with GM1 [14C]-labelled at the sialic acid acetyl group |
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Authors: | Vanna Chigorno Marina Pitto Gloria Cardace Domenico Acquotti Günther Kirschner Sandro Sonnino Riccardo Ghidoni Guido Tettamanti |
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Institution: | (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;(2) Department of Chemistry, Fidia Research Laboratories, Abano Terme, Italy |
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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.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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Keywords: | labelled G
M1
-ganglioside N-[
14
C]-acetylneuraminic acid cultured fibroblasts |
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