Incorporation of N-acetylglucosamine into endogenous acceptors of rough microsomes from rat liver: stimulation by GTP after treatment with pyrophosphate.
- 1 March 1977
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 74 (3), 1095-1099
- https://doi.org/10.1073/pnas.74.3.1095
Abstract
On incubation of UDP-N-acetyl[14C]glucosamine [UDP-GlcNAc] and GDP-mannose with rough microsomes from rat liver, GlcNAc incorporation occurred mostly into acid-labile products extractable by chloroform/methanol that were presumed to be dolichol pyrophosphate GlcNAc and, for a small part, dolichol pyrophosphate N,N''-diacetylchitobiose [DPD]. The production of this latter glycolipid was greatly enhanced when rough microsomes, previously treated with pyrophosphate, were incubated in the presence of GTP. Under these particular conditions, high levels of radioactivity were obtained in products precipitated by trichloroacetic acid and not extracted by chloroform/methanol. The label appeared in acid-labile and in acid-resistant products, presumed to be dolichol pyrophosphate mannosylated oligosaccharides, and glycoproteins, respectively. From these results, DPD appears to be an obligate metabolic intermediate in protein glycosylation by this system. Whether or not DPD must be mannosylated prior to transfer of the N,N''-diacetylchitobiose moiety to protein is an open matter, because GlcNAc was also incorporated into acid-resistant products when the reaction was carried out in absence of GDP-mannose.This publication has 19 references indexed in Scilit:
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