Fluorescent derivatives of ganglioside GM1 function as receptors for cholera toxin.
A fluorescent derivative of ganglioside GM1 was prepared by oxidation of the sialic acid residue with sodium periodate and reaction of the resulting aldehyde with Lucifer yellow CH. The biological activity of the fluorescent derivative was compared with that of native GM1 using GM1-deficient rat glioma C6 cells. When the cells were exposed to either native or fluorescent GM1, their ability to bind 125I-labeled cholera toxin was increased to a similar extent. This increase in binding was directly proportional to the amount of ganglioside added to the medium. The affinity of the toxin for cells treated with either native or fluorescent GM1 also was similar. More importantly, the fluorescent GM1 was as effective as native GM1 in enhancing the responsiveness of the cells to cholera toxin. Thus, the ganglioside-treated cells exhibited a 9-fold increase in toxin-stimulated cyclic AMP production over cells not exposed to GM1. There was a similar increase in iodotoxin binding and toxin-stimulated cyclic AMP accumulation in cells treated with other GM1 derivatives containing rhodaminyl or dinitrophenyl groups. On the basis of these results, it is clear that these modified gangliosides retain the ability to function as receptors for cholera toxin. Consequently, fluorescent gangliosides are likely to be useful as probes for investigating the dynamics and function of these membrane components.
D000818:Animals; D002460:Cell Line; D002772:Cholera Toxin / Q000378:metabolism*; D002855:Chromatography, Thin Layer; D000242:Cyclic AMP / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism*; D005677:G(M1) Ganglioside / Q000378:metabolism*; D005910:Glioma / Q000378:metabolism*; D007546:Isoquinolines / Q000378:metabolism*; D007700:Kinetics; D051381:Rats; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic / Q000378:metabolism*
D005456:Fluorescent Dyes; D007546:Isoquinolines; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; C023552:choleragen receptor; D005677:G(M1) Ganglioside; D002772:Cholera Toxin; C017475:lucifer yellow; D000242:Cyclic AMP