Non-Radioactive Analysis of Lipid-Linked Oligosaccharide Compositions by Fluorophore-Assisted Carbohydrate Electrophoresis

Ningguo Gao, Mark A. Lehrman

Research output: Contribution to journalArticlepeer-review

33 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)3-20
Number of pages18
JournalMethods in Enzymology
Volume415
DOIs
StatePublished - 2006

ASJC Scopus subject areas

  • Biochemistry

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