Simultaneous determination of nucleotide sugars with ion-pair reversed-phase HPLC

Kazuki Nakajima, Shinobu Kitazume, Takashi Angata, Reiko Fujinawa, Kazuaki Ohtsubo, Eiji Miyoshi, Naoyuki Taniguchi

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

Nucleotide sugars are important in determining cell surface glycoprotein glycosylation, which can modulate cellular properties such as growth and arrest. We have developed a conventional HPLC method for simultaneous determination of nucleotide sugars. A mixture of nucleotide sugars (CMP-NeuAc, UDP-Gal, UDP-Glc, UDP-GalNAc, UDP- GlcNAc, GDP-Man, GDP-Fuc and UDP-GlcUA) and relevant nucleotides were perfectly separated in an optimized ion-pair reversed-phase mode using Inertsil ODS-4 and ODS-3 columns. The newly developed method enabled us to determine the nucleotide sugars in cellular extracts from 1×106 cells in a single run. We applied this method to characterize nucleotide sugar levels in breast and pancreatic cancer cell lines and revealed that the abundance of UDP-GlcNAc, UDP-GalNAc, UDP-GlcUA and GDP-Fuc were a cell-type-specific feature. To determine the physiological significance of changes in nucleotide sugar levels, we analyzed their changes by glucose deprivation and found that the determination of nucleotide sugar levels provided us with valuable information with respect to studying the overview of cellular glycosylation status.

Original languageEnglish
Pages (from-to)865-871
Number of pages7
JournalGlycobiology
Volume20
Issue number7
DOIs
Publication statusPublished - 01-01-2010

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Uridine Diphosphate
Sugars
Nucleotides
High Pressure Liquid Chromatography
Ions
Glycosylation
Cytidine Monophosphate
Membrane Glycoproteins
Cell Extracts
Pancreatic Neoplasms
Cells
Breast Neoplasms
Glucose
Cell Line
Growth

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Nakajima, K., Kitazume, S., Angata, T., Fujinawa, R., Ohtsubo, K., Miyoshi, E., & Taniguchi, N. (2010). Simultaneous determination of nucleotide sugars with ion-pair reversed-phase HPLC. Glycobiology, 20(7), 865-871. https://doi.org/10.1093/glycob/cwq044
Nakajima, Kazuki ; Kitazume, Shinobu ; Angata, Takashi ; Fujinawa, Reiko ; Ohtsubo, Kazuaki ; Miyoshi, Eiji ; Taniguchi, Naoyuki. / Simultaneous determination of nucleotide sugars with ion-pair reversed-phase HPLC. In: Glycobiology. 2010 ; Vol. 20, No. 7. pp. 865-871.
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Nakajima, K, Kitazume, S, Angata, T, Fujinawa, R, Ohtsubo, K, Miyoshi, E & Taniguchi, N 2010, 'Simultaneous determination of nucleotide sugars with ion-pair reversed-phase HPLC', Glycobiology, vol. 20, no. 7, pp. 865-871. https://doi.org/10.1093/glycob/cwq044

Simultaneous determination of nucleotide sugars with ion-pair reversed-phase HPLC. / Nakajima, Kazuki; Kitazume, Shinobu; Angata, Takashi; Fujinawa, Reiko; Ohtsubo, Kazuaki; Miyoshi, Eiji; Taniguchi, Naoyuki.

In: Glycobiology, Vol. 20, No. 7, 01.01.2010, p. 865-871.

Research output: Contribution to journalArticle

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Nakajima K, Kitazume S, Angata T, Fujinawa R, Ohtsubo K, Miyoshi E et al. Simultaneous determination of nucleotide sugars with ion-pair reversed-phase HPLC. Glycobiology. 2010 Jan 1;20(7):865-871. https://doi.org/10.1093/glycob/cwq044