RRC ID 66851
Author Fujitani N, Ariki S, Hasegawa Y, Uehara Y, Saito A, Takahashi M.
Title Integrated Structural Analysis of N-Glycans and Free Oligosaccharides Allows for a Quantitative Evaluation of ER Stress.
Journal Biochemistry
Abstract Endoplasmic reticulum (ER) stress has been reported in a variety of diseases. Although ER stress can be detected using specific markers, it is still difficult to quantitatively evaluate the degree of stress and to identify the cause of the stress. The ER is the primary site for folding of secretory or transmembrane proteins as well as the site where glycosylation is initiated. This study therefore postulates that tracing the biosynthetic pathway of asparagine-linked glycans (N-glycans) would be a reporter for reflecting the state of the ER and serve as a quantitative descriptor of ER stress. Glycoblotting-assisted mass spectrometric analysis of the HeLa cell line enabled quantitative determination of the changes in the structures of N-glycans and degraded free oligosaccharides (fOSs) in response to tunicamycin- or thapsigargin-induced ER stress. The integrated analysis of neutral and sialylated N-glycans and fOSs showed the potential to elucidate the cause of ER stress, which cannot be readily done by protein markers alone. Changes in the total amount of glycans, increase in the ratio of high-mannose type N-glycans, increase in fOSs, and changes in the ratio of sialylated N-glycans in response to ER stress were shown to be potential descriptors of ER stress. Additionally, drastic clearance of accumulated N-glycans was observed in thapsigargin-treated cells, which may suggest the observation of ER stress-mediated autophagy or ER-phagy in terms of glycomics. Quantitative analysis of N-glycoforms composed of N-glycans and fOSs provides the dynamic indicators reflecting the ER status and the promising strategies for quantitative evaluation of ER stress.
Volume 60(21)
Pages 1708-1721
Published 2021-6-1
DOI 10.1021/acs.biochem.0c00969
PMID 33983715
MeSH Asparagine / metabolism Biomarkers Endoplasmic Reticulum / pathology* Endoplasmic Reticulum Stress / physiology* Glycosylation HeLa Cells Humans Mannose / metabolism Mass Spectrometry / methods Membrane Proteins / metabolism Oligosaccharides / chemistry Oligosaccharides / metabolism Polysaccharides / chemistry Polysaccharides / metabolism Structure-Activity Relationship Tunicamycin / pharmacology
IF 2.865
Human and Animal Cells HeLa