Localization of galactosyl- and sialyltransferase by immunofluorescence: evidence for different sites.
Polyclonal rabbit antisera against soluble human milk galactosyltransferase and bovine colostrum sialyltransferase were used to localize by indirect immunofluorescence the respective intracellular enzymes in primary cultures from bovine fetal kidneys and established cell lines of human and bovine fibroblasts. Staining for galactosyltransferase was juxtanuclear and crescent shaped in epitheloid cells; a similar staining, occasionally perinuclear and sparsely distributed in the cytoplasm, was found in fibroblasts. In contrast, staining for sialyltransferase in epitheloid kidney cells derived from the same primary culture was observed predominantly in cytoplasmic vesicles that were spread over the whole cytoplasm. Sialyltransferase-positive vesicles had a similar distribution in fibroblasts and often appeared concentrated around an unstained Golgi area. Thus, in both cell types galactosyl- and sialyltransferase were localized in different subcellular compartments. Since both galactosyl- and sialyltransferase participate in formation of the terminal glycan NeuAc(alpha 2----6)Gal(beta 1----4)GlcNAc(Neu, neuraminic acid) present in many N-glycosidic complex types of glycans, different subcellular compartments for these enzymes support a model of functional compartmentalization of the Golgi apparatus that is compatible with an assembly-line model for glycan chain elongation and termination.
D000818:Animals; D000936:Antigen-Antibody Complex; D002417:Cattle; D003126:Colostrum / Q000201:enzymology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology; D005455:Fluorescent Antibody Technique; D005700:Galactosyltransferases / Q000032:analysis*; D006801:Humans; D007106:Immune Sera; D007668:Kidney / Q000166:cytology / Q000201:enzymology; D008895:Milk, Human / Q000201:enzymology; D011247:Pregnancy; D012799:Sialyltransferases / Q000032:analysis*; D014166:Transferases / Q000032:analysis*
D000936:Antigen-Antibody Complex; D007106:Immune Sera; D014166:Transferases; D005700:Galactosyltransferases; D012799:Sialyltransferases