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 journalArticlepeer-review

116 Citations (Scopus)


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
Issue number7
Publication statusPublished - 07-2010

All Science Journal Classification (ASJC) codes

  • Biochemistry


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