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Glycobiology Advance Access published online on March 23, 2007
Glycobiology, doi:10.1093/glycob/cwm029
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© The Author 2007. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Review

Teaching Dolichol-Linked Oligosaccharides More Tricks With Alternatives To Metabolic Radiolabeling

Mark A. Lehrman

Dept. Pharmacology, University of Texas Southwestern Medical Center, 6001 Forest Park Rd. Dallas, TX 75390-9041

Tel 214-645-6172 Fax 214-645-6173 email mark.lehrman{at}utsouthwestern.edu

Received on February 20, 2007; revised on February 20, 2007; accepted on March 7, 2007

The dolichol cycle involves synthesis of the lipid-linked oligosaccharide (LLO) Glc3Man9GlcNAc2-P-P-dolichol (G3M9Gn2-P-P-Dol), transfer of G3M9Gn2 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 [3H]-labeled or [14C]-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.

Key words: lipid-linked oligosaccharide / dolichol / N-linked glycosylation / fluorophore-assisted carbohydrate electrophoresis


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N. Gao, J. Shang, and M. A Lehrman
Unexpected Basis for Impaired Glc3Man9GlcNAc2-P-P-Dolichol Biosynthesis by Elevated Expression of GlcNAc-1-P Transferase
Glycobiology, January 1, 2008; 18(1): 125 - 134.
[Abstract] [Full Text] [PDF]


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