Cerebroside sulfotransferase in Golgi-enriched fractions from rat brain.
Golgi-enriched fractions were prepared from brainstems of 17-day-old rats by first floating off myelin, then fractionating the remaining pellet by a series of differential and density gradient centrifugations in sucrose. Fractions enriched in Golgi membranes were recovered at 0.46/0.76 M and 0.76/0.87 M interfaces on the final sucrose gradient as indicated by morphology and the biochemical markers thiamine pyrophosphatase and [3H]fucose-labeled glycoprotein. Morphology of the two fractions indicated very little contamination with myelin lamellae; however, the presence of significant levels of 2',3'-cyclic nucleotidase in the lighter fraction suggested a contribution from oligodendroglial or myelin-related membranes. Cerebroside sulfotransferase was highly enriched in the lighter Golgi-enriched fraction relative to the denser fraction, the post-34,880 X g microsomes, and the myelin-like fraction. In contrast, ceramide galactosyl transferase was more evenly distributed among the fractions. Our results show a more highly localized distribution of sulfatide synthesis than of galactocerebroside synthesis, probably in Golgi membranes or oligodendroglia-related membranes with similar properties.
D000818:Animals; D001921:Brain / Q000201:enzymology*; D002499:Centrifugation, Density Gradient; D002554:Cerebrosides / Q000096:biosynthesis; D006056:Golgi Apparatus / Q000201:enzymology*; D008854:Microscopy, Electron; D009186:Myelin Sheath / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D013347:Subcellular Fractions / Q000648:ultrastructure; D013433:Sulfoglycosphingolipids / Q000096:biosynthesis; D015238:Sulfotransferases; D013466:Sulfurtransferases / Q000378:metabolism*
D002554:Cerebrosides; D013433:Sulfoglycosphingolipids; D013466:Sulfurtransferases; D015238:Sulfotransferases; C020556:galactosylceramide sulfotransferase