期刊
GLYCOBIOLOGY
卷 17, 期 8, 页码 75R-85R出版社
OXFORD UNIV PRESS INC
DOI: 10.1093/glycob/cwm029
关键词
dolichol; fluorophore-assisted carbohydrate electrophoresis; lipid-linked oligosaccharide; N-linked glycosylation
资金
- NIDDK NIH HHS [DK70954] Funding Source: Medline
- NIGMS NIH HHS [R01 GM038545-20, GM38545, R01 GM038545] Funding Source: Medline
The dolichol cycle involves synthesis of the lipid-linked oligosaccharide (LLO) GlC(3)Man(9)GlcNAC(2)-P-P-dolichol (G(3)M(9)Gn(2)-P-P-Dol), transfer of G(3)M(9)Gn(2) to asparaginyl residues of nascent endoplasmic reticulum (ER) polypeptides by oligosaccharyltransferase (OT), and recycling of the resultant Dol-P-P to Dol-P for new rounds of LLO synthesis. The importance of the dolichol cycle in secretory and membrane protein biosynthesis, ER function, and human genetic disease is now widely accepted. Elucidation of the fundamental properties of the dolichol cycle in intact cells was achieved through the use of radioactive sugar precursors, typically 13 HI-labeled or [C-14]-labeled D-mannose, D-galactose, or D-glucosamine. However, difficulties were encountered with cells or tissues not amenable to metabolic labeling, or in experiments influenced by isotope dilution, variable rates of LLO turnover, or special culture conditions required for the use of radioactive sugars. This article will review recently developed alternatives for LLO analysis that do not rely upon metabolic labeling with radioactive precursors, and thereby circumvent these problems. New information revealed by these methods with regard to regulation, genetic disorders, and evolution of the dolichol cycle, as well as caveats of radiolabeling techniques, will be discussed.
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