Non-radioactive analysis of lipid-linked oligosaccharide compositions by fluorophore-assisted carbohydrate electrophoresis - PubMed (original) (raw)

Non-radioactive analysis of lipid-linked oligosaccharide compositions by fluorophore-assisted carbohydrate electrophoresis

Ningguo Gao et al. Methods Enzymol. 2006.

Abstract

Lipid-linked oligosaccharides (LLOs) are the donors of glycans that modify newly synthesized proteins in the endoplasmic reticulum (ER) of eukaryotes, resulting in formation of N-linked glycoproteins. The vast majority of LLO analyses have relied on metabolic labeling with radioactive sugar precursors, but these approaches have technical limitations resulting in many important questions about LLO synthesis being left unanswered. Here we describe the application of a facile non-radioactive technique, fluorophore-assisted carbohydrate electrophoresis (FACE), which circumvents these limitations. With FACE, steady-state LLO compositions can be determined quantitatively from cell cultures and animal tissues. We also present FACE methods for analysis of phosphosugars and nucleotide sugars, which are metabolic precursors of LLOs.

PubMed Disclaimer

Cited by

Regulation of protein glycosylation and sorting by the Golgi matrix proteins GRASP55/65.
Xiang Y, Zhang X, Nix DB, Katoh T, Aoki K, Tiemeyer M, Wang Y. Xiang Y, et al. Nat Commun. 2013;4:1659. doi: 10.1038/ncomms2669. Nat Commun. 2013. PMID: 23552074 Free PMC article.
A zebrafish model of congenital disorders of glycosylation with phosphomannose isomerase deficiency reveals an early opportunity for corrective mannose supplementation.
Chu J, Mir A, Gao N, Rosa S, Monson C, Sharma V, Steet R, Freeze HH, Lehrman MA, Sadler KC. Chu J, et al. Dis Model Mech. 2013 Jan;6(1):95-105. doi: 10.1242/dmm.010116. Epub 2012 Aug 16. Dis Model Mech. 2013. PMID: 22899857 Free PMC article.
Use of a Glycan Library Reveals a New Model for Enteric Virus Oligosaccharide Binding and Virion Stabilization.
Lu H, Lehrman MA, Pfeiffer JK. Lu H, et al. J Virol. 2020 Feb 28;94(6):e01894-19. doi: 10.1128/JVI.01894-19. Print 2020 Feb 28. J Virol. 2020. PMID: 31852778 Free PMC article.
Mannose-6-phosphate regulates destruction of lipid-linked oligosaccharides.
Gao N, Shang J, Huynh D, Manthati VL, Arias C, Harding HP, Kaufman RJ, Mohr I, Ron D, Falck JR, Lehrman MA. Gao N, et al. Mol Biol Cell. 2011 Sep;22(17):2994-3009. doi: 10.1091/mbc.E11-04-0286. Epub 2011 Jul 7. Mol Biol Cell. 2011. PMID: 21737679 Free PMC article.
Nogo-B receptor is necessary for cellular dolichol biosynthesis and protein N-glycosylation.
Harrison KD, Park EJ, Gao N, Kuo A, Rush JS, Waechter CJ, Lehrman MA, Sessa WC. Harrison KD, et al. EMBO J. 2011 May 13;30(12):2490-500. doi: 10.1038/emboj.2011.147. EMBO J. 2011. PMID: 21572394 Free PMC article.

Non-radioactive analysis of lipid-linked oligosaccharide compositions by fluorophore-assisted carbohydrate electrophoresis - PubMed (original) (raw)