Xylosylation and glucuronosylation reactions in rat liver Golgi apparatus and endoplasmic reticulum.
We have studied in rat liver the subcellular sites and topography of xylosylation and galactosylation reactions occurring in the biosynthesis of the D-glucuronic acid-galactose-galactose-D-xylose linkage region of proteoglycans and of glucuronosylation reactions involved in both glycosaminoglycan biosynthesis and bile acid and bilirubin conjugation. The specific translocation rate of UDP-xylose into sealed, "right-side-out" vesicles from the Golgi apparatus was 2-5-fold higher than into sealed right-side-out vesicles from the rough endoplasmic reticulum (RER). Using the above vesicle preparations, we only detected endogenous acceptors for xylosylation in the Golgi apparatus-rich fraction. The specific activity of xylosyltransferase (using silk fibroin as exogenous acceptor) was 50-100-fold higher in Golgi apparatus membranes than in those from the RER. Previous studies had shown that UDP-galactose is translocated solely into vesicles from the Golgi apparatus. In these studies, we found the specific activity of galactosyltransferase I to be 40-140-fold higher in membranes from the Golgi apparatus than in those from the RER. The specific translocation rate of UDP-D-glucuronic acid into vesicles from the Golgi apparatus was 10-fold higher than into those from the RER, whereas the specific activity of glucuronosyltransferase (using chondroitin nonasaccharide as exogenous acceptor) was 12-30-fold higher in Golgi apparatus membranes than in those from the RER. Together, the above results strongly suggest that, in rat liver, the biosynthesis of the above-described proteoglycan linkage region occurs in the Golgi apparatus. The specific activity of glucuronosyltransferase, using bile acids and bilirubin as exogenous acceptor, was 10-25-fold higher in RER membranes than those from the Golgi apparatus. This suggests that transport of UDP-D-glucuronic acid into the RER lumen is not required for such reactions.
D000818:Animals; D002236:Carbohydrate Conformation; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005690:Galactose / Q000378:metabolism; D005965:Glucuronates / Q000378:metabolism*; D020723:Glucuronic Acid; D014453:Glucuronosyltransferase / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000648:ultrastructure*; D051381:Rats; D014531:Uridine Diphosphate Galactose / Q000378:metabolism; D014540:Uridine Diphosphate Xylose / Q000378:metabolism; D014994:Xylose / Q000378:metabolism*
D005965:Glucuronates; D014531:Uridine Diphosphate Galactose; D014540:Uridine Diphosphate Xylose; D020723:Glucuronic Acid; D014994:Xylose; D014453:Glucuronosyltransferase; D005690:Galactose