Stimulation of lipid-linked oligosaccharide assembly during oviduct differentiation

• Published in: The Journal of biological chemistry Vol. 258; no. 24; pp. 15095 - 15100
• Main Authors: Hayes, G R, Lucas, J J
• Format: Journal Article
• Language: English
• Published: Bethesda, MD Elsevier Inc 25-12-1983; American Society for Biochemistry and Molecular Biology
• Subjects: Animals; Biological and medical sciences; Cell Differentiation - drug effects; Chickens; Diethylstilbestrol - pharmacology; Dolichol Phosphates - metabolism; Embryology: invertebrates and vertebrates. Teratology; Female; Fundamental and applied biological sciences. Psychology; Guanosine Diphosphate Mannose - metabolism; Kinetics; Lipid Metabolism; Mannose - metabolism; Organogenesis. Physiological fonctions; Oviducts - cytology; Polyisoprenyl Phosphate Oligosaccharides - biosynthesis; Polyisoprenyl Phosphate Sugars - biosynthesis
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(17)43776-8

Regulation of dolichyl phosphate-linked oligosaccharide assembly has been studied during the course of diethylstilbestrol-induced chick oviduct differentiation. Oviduct membranes from treated chicks form 4.6 times as much GlcNAc-P-P-Dol and GlcNAc2-P-P-Dol upon incubation with UDP-[14C]GlcNAc and MgCl2 than do membranes from untreated chicks. Assembly of oligosaccharide-lipid was studied by incubation of membranes with purified exogenous [14C]GlcNAc2-P-P-Dol and GDP-Man. Man transfer required a divalent cation (10 mM Mg2+) and detergent (0.5% Nonidet P-40 is optimal) and occurs in the presence of amphomycin (500 micrograms/ml). The apparent Km for GDP-Man is 1 microM and for [14C]GlcNAc2-P-P-Dol is 0.45 microM. The products are a series of sequentially formed dolichyl pyrophosphate-linked saccharides up to Man5GlcNAc2, the first of which is Man beta 1,4GlcNAc2. The same products are formed either in the presence or absence of amphomycin. Conversion of GlcNAc2-P-P-Dol to higher oligosaccharides is stimulated 3-fold by estrogen treatment of chicks. Similarly, the conversion of partially purified exogenously added Man beta-[14C]GlcNAc2-P-P-Dol is 4.6-fold higher after diethylstilbestrol treatment.