High-speed and high-sensitive methods of carbohydrates based on mass isotopomer analysis with the aim of monitoring glycan dynamics

2014 Fiscal Year Final Research Report

• Project/Area Number: 24790301
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: General medical chemistry
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: NAKAJIMA Kazuki 独立行政法人理化学研究所, 脳科学総合研究センター, 研究員 (10442998)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: 糖鎖修飾 / 糖ヌクレオチド / 質量分析 / クロマトグラフィー / グルコース代謝 / シアル酸

Outline of Final Research Achievements

In order to clarify biological significance of glycan dynamics, we have developed a tracing method for UDP-GlcNAc synthesis and utilization, and GlcNAc utilization using 13C6-glucose and 13C2-glucosamine, respectively, followed by the analysis of mass isotopomers. Because nucleotide sugars an important building block in glycan biosynthesis. Analysis of isotopomers of UDP-HexNAc and CMP-NeuAc, labeled with 13C6-glucose, revealed the relative contributions of metabolic pathways leading to UDP-GlcNAc synthesis and utilization. Using 13C2-glucosamine, the diversity of glycan turnover was observed in each glycan. In pancreatic insulinoma cells, the labeling efficiency of CMP-NeuAc was lower compared with that in hepatoma cells, indicating that metabolic flows are responsible for the low sialylation in the insulinoma cells. Thus, our strategy should be useful for systematically tracing each stage of cellular glycan dynamics.

Free Research Field

生化学