Synthesis of retinylphosphate mannose in yeast and its possible involvement in lipid-linked oligosaccharide formation.
A membrane fraction from Saccharomyces cerevisiae as well as a mannosyltransferase purified therefrom was shown to catalyze the transfer of mannose from GDPmannose to retinyl phosphate. The product formed has chromatographic and chemical properties characteristic for retinylphosphate mannose. The enzyme requires divalent cations. Mg2+ is more effective than Mn2+ with an optimum concentration around 25 mM. Amphomycin at a concentration of 0.1 mg/ml inhibits the reaction to 50%. Glycosyl transfer was specific for mannose residues from GDPmannose and did not occur with dolichylphosphate mannose nor with UDP galactose; UDPglucose is a poor donor. Formation of retinylphosphate mannose is inhibited by dolichyl phosphate. This observation as well as similarities between retinylphosphate mannose and dolichylphosphate mannose synthesis in respect to ion requirement, inhibition by amphomycin are suggestive that both reactions are catalyzed by one and the same enzyme. In experiments studying the glycosyl donor specificity in the assembly of lipid-linked oligosaccharide intermediates involved in N-glycosylation of proteins, it could be demonstrated that retinylphosphate mannose can replace dolichylphosphate mannose in the final steps of mannosylation.
D002384:Catalysis; D004224:Diterpenes; D004288:Dolichol Phosphates / Q000494:pharmacology; D006155:Guanosine Diphosphate Mannose / Q000378:metabolism; D050356:Lipid Metabolism; D009844:Oligosaccharides / Q000096:biosynthesis*; D011103:Polyisoprenyl Phosphate Monosaccharides / Q000096:biosynthesis* / Q000502:physiology; D011105:Polyisoprenyl Phosphate Sugars / Q000096:biosynthesis*; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D013379:Substrate Specificity
D004224:Diterpenes; D004288:Dolichol Phosphates; D009844:Oligosaccharides; D011103:Polyisoprenyl Phosphate Monosaccharides; D011105:Polyisoprenyl Phosphate Sugars; C028306:dolichol monophosphate; D006155:Guanosine Diphosphate Mannose; C013101:mannosylretinylphosphate