1. Two-step derivatization and mass spectral distinction of α2,3 and α2,6 sialic acid linkages on N-glycans by MALDI-TOF
- Author
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Zengqi Tan, Ganglong Yang, Feng Guan, Shang Yang, and Xiaoman Zhou
- Subjects
Glycan ,Glycosylation ,Chromatography ,biology ,Chemistry ,02 engineering and technology ,General Chemistry ,010402 general chemistry ,021001 nanoscience & nanotechnology ,Mass spectrometry ,01 natural sciences ,Fetuin ,0104 chemical sciences ,Sialic acid ,carbohydrates (lipids) ,chemistry.chemical_compound ,Ammonium hydroxide ,biology.protein ,0210 nano-technology ,Derivatization ,Dimethylamine - Abstract
Sialylation is one of important glycosylation in human beings and plays an important role in cancer development. α2,3-Linked and α2,6-linked sialic acids are normally observed on the end of N-glycans and have different functions. Derivatization on sialic acid was designed to detect the different linkages by MALDI-TOF MS. In this study, a two-step derivatization by dimethylamine and ammonium hydroxide was improved to modify the sialic acid and made it easier to detect the different linkages of sialic acids on MALDI-TOF MS. Using this derivatization method, specific sialic acids linkages on N-glycans of protein samples such as fetuin and lactoferrin were detected. For complex cell samples, increased α2,3-linked and α2,6-linked sialic acids on bi-antennary and tri-antennary N-glycans were observed in A549 cells induced by hypoxia environment. Taken together, our two-step derivatization of sialic acids offers a simple and accurate way to detect specific linkages on N-glycans with MALDI-TOF mass spectrometer.
- Published
- 2019