Conversion of short-chain ceramides to short-chain ceramide GM3 in B16 melanoma cells

We report that short-chain ceramide (Cer), C2- and C6-Cer, were immediately glycosylated and finally converted to short-chain Cer GM3 in B16 melanoma cells. By addition of either C2- or C6-Cer to a cell culture of B16 melanoma in the presence of [14C]Gal, the radiolabeled precursor, was incorporated into each of two novel glycosphingolipids (GSLs) within 30 min along with synthesis of normal GSLs. These novel GSLs were identified as C2-, C6-Cer cerebrosides and C2-, C6-Cer GM3, respectively. In comparison with C2-Cer, C6-Cer was found to be much more efficiently converted to the GSLs, whereas no glycosylated sphingosine was detectable when it was added in place of short-chain Cer.