김진일 (국립창원대학교)

임재민 (국립창원대학교)

임재민 (국립창원대학교)

김진일 (국립창원대학교)

신도윤 (국립창원대학교)

김도현 (국립창원대학교)

N-glycosylation is one of the most important post-translation modifications. Glycans formed through it can affect biochemical properties and physiological functions of in vivo. In order to decode the correlation of glycan’s prevalence with their physiological contribution, many mass spectrometry (MS) with stable isotope labeling-based methods have been developed for the relative quantification of glycans. Based on the MILPIG (metabolic isotope labeling of polysaccharides with isotopic glucose), we establish an isotope labeling method and extend the quantitative glycomics using C. elegans as the model system. The main prey of C. elegans is cornmeal, peptone or E.coil. Oligidic medium containing these was prepared from normal medium for the culture of C. elegans. However, these cannot be applied to the MILPIG strategy, where glucose must be strictly controlled, because the glucose content is unknown.

To apply the MILPIG strategy to the C. elegans model, holidic medium capable of controlling glucose concentration was prepared in glucose-free medium without cornmeal, peptone or E. coil, which cannot control glucose concentration. By comparing the growth rates of C. elegans cultured in the oligidic and holidic media, we applied the MILPIG strategy to the C. elegans model cultured in holidic media.

Afterwards, C. elegans was grown in glucose concentration-controlled medium, the glycans were purified, and the mass spectrum of C. elegans’s glycans were obtained by mass spectrometry. Additionally, mass spectrum of isotopically labeled glycans were obtained in media containing isotopically labeled glucose (1-¹³C₁) instead of normal glucose. Quantitative information was provided through comparison of the area of isotope clusters in mass spectrum from the same amount of normal and isotopically labeled glycans.