Glucuronosyl-N-acetylglucosaminyl pyrophosphoryldolichol. Formation in SV40-transformed human lung fibroblasts and biosynthesis in rat lung microsomal preparations.
Incubation of SV40-transformed human lung fibroblasts with [3H]glucosamine for 1 h. followed by chloroform:methanol extraction and thin layer chromatographic analysis, revealed the presence of a major radioactive lipid that was isolated and characterized as GIcUA-(1 leads to 4)-GlcNAc-P-P-dolichol. An identical lipid was formed in smaller quantities under similar incubation conditions in several fibroblastic lines, HeLa cells, and in mouse L cells. Rat lung microsomal preparations catalyze the synthesis of the disaccharide lipid in the following sequence of reactions: UDP-[3H]GlcNAc + dolichol-P leads to [3H]GlcNAc-P-P-dolichol (1) [3H]GlcNAc-P-P-dolichol + UDP-[14C]GlcUA leads to [14C]GlcUA-[3H]GlcNAc-P-P-dolichol (2) The double-labeled lipid was identical to the lipid isolated from SV40-transformed fibroblasts with regard to its behavior on thin layer and silicic acid chromatography. Further, the double-labeled disaccharide released from the lipid by mild acid hydrolysis was identical to GlcUA-(1 leads to 4)-GlcNAc in its chromatographic and electrophoretic behavior and in its composition. The occurrence of a polyprenol derivative of GlcUA-(1 leads to 4)-GlcNAc suggests a possible role for this lipid in the biosynthesis of the repeating disaccharide units of proteoglycans, such as heparin.
D000818:Animals; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D004224:Diterpenes / Q000096:biosynthesis*; D004286:Dolichols / Q000031:analogs & derivatives / Q000096:biosynthesis*; D005347:Fibroblasts / Q000378:metabolism; D005944:Glucosamine / Q000378:metabolism; D005958:Glucosephosphates / Q000096:biosynthesis*; D006801:Humans; D008055:Lipids / Q000096:biosynthesis; D008168:Lung / Q000378:metabolism*; D008297:Male; D008861:Microsomes / Q000378:metabolism; D011104:Polyisoprenyl Phosphate Oligosaccharides; D014176:Protein Biosynthesis; D051381:Rats; D013539:Simian virus 40
D004224:Diterpenes; D004286:Dolichols; D005958:Glucosephosphates; D008055:Lipids; D011104:Polyisoprenyl Phosphate Oligosaccharides; C014572:glucuronosyl-N-acetylglucosaminyl pyrophosphoryldolichol; D005944:Glucosamine