Golgi galactosyltransferase contains serine-linked phosphate.

In HeLa and HepG2 cells the Golgi complex enzyme galactosyltransferase became phosphorylated following incubation with 32Pi-Analysis on sodium dodecyl sulphate/polyacrylamide gel electrophoresis revealed incorporation of 32P into the mature 54-kDa form. This phosphorylation was independent of protein synthesis. Serine was identified as the sole phosphorylated amino acid; no radioactive phosphate was detected on N-linked oligosaccharide. The phosphate-labelled galactosyltransferase has the same turnover as [35S]methionine-labelled polypeptides (t1/2 = 20 h). Soluble enzyme, released by the cells, contained very little phosphate relative to that which remained cell-associated. Charge heterogeneity arising from phosphorylation contributes in part to the heterodispersed appearance of the enzyme on two-dimensional gels, as the degree of radioactive phosphate differs among the different iso-enzymes.