Abstract
Bovine aortic smooth-muscle cells, bovine aortic endothelial cells, and IMR-90 human embryonic lung fibroblasts were tested to determine their ability to use cysteine or cysteine metabolites as a source of sulphate (SO4). Cells were incubated in SO4-depleted medium containing [3H]glucosamine plus 0.2 mM-cystine, 0.3 mM-cysteinesulphinic acid or 0.3 mM-sulphite (SO3). The [3H]chondroitin sulphate produced by the different cells was found to vary considerably in degree of sulphation under these conditions. One line of smooth-muscle cells utilized cysteine effectively as a SO4 source and thus produced chondroitin sulphate which was highly sulphated. IMR-90 fibroblasts produced partly sulphated chondroitin sulphate under these conditions, while another smooth-muscle cell line could not utilize cysteine, but could utilize cysteinesulphinic acid as a partial SO4 source. In contrast with the above cells, endothelial cells could not use cysteine or cysteinesulphinic acid as a source of SO4 and produced chondroitin with almost no SO4. All of the cells were able to utilize SO3. Incubation of the cells in the SO4-depleted medium containing [35S]cysteine confirmed that only the first line of smooth-muscle cells could convert significant amounts of [35S]cysteine to 35SO4. Furthermore, the addition of 0.4 mM inorganic SO4 did not inhibit the production of SO4 from cysteine by these cells.