Mapping the distribution of Golgi enzymes involved in the construction of complex oligosaccharides.
The distribution of beta 1,2 N-acetylglucosaminyltransferase I (NAGT I), alpha 1,3-1,6 mannosidase II (Mann II), beta 1,4 galactosyltransferase (GalT), alpha 2,6 sialyltransferase (SialylT) was determined by immuno-labelling of cryo-sections from HeLa cell lines. Antibody labelling in the HeLa cell line was made possible by stable expression of epitope-tagged forms of these proteins or forms from species to which specific antibodies were available. NAGT I and Mann II had the same distribution occupying the medial and trans cisternae of the stack. GalT and SialylT also had the same distribution but they occupied the trans cisterna and the trans-Golgi network (TGN). These results generalise our earlier observations on the overlapping distribution of Golgi enzymes and show that each of the trans compartments of the Golgi apparatus in HeLa cells contains unique mixtures of those Golgi enzymes involved in the construction of complex, N-linked oligosaccharides.
Rabouille
C
C
Cell Biology Laboratory, Imperial Cancer Research Fund, London, UK.
Hui
N
N
Hunte
F
F
Kieckbusch
R
R
Berger
E G
EG
Warren
G
G
Nilsson
T
T
D000818:Animals; D000906:Antibodies; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D017931:DNA Primers; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000201:enzymology* / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000032:analysis*; D009844:Oligosaccharides / Q000096:biosynthesis*; D016133:Polymerase Chain Reaction; D051381:Rats; D012799:Sialyltransferases / Q000032:analysis*; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D000906:Antibodies; D017931:DNA Primers; D009844:Oligosaccharides; D017351:N-Acetylglucosaminyltransferases; C024850:alpha-1,3-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I; D012799:Sialyltransferases; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase