Abstract
BACKGROUND Mass spectrometry-based proteomics and glycomics reveal post-translational modifications providing significant biological insights beyond the scope of genomic sequencing.
AIM To characterize the N-linked glycoproteomic profile in esophageal squamous cell carcinoma (ESCC) via two complementary approaches.
METHODS Using tandem multilectin affinity chromatography for enrichment of N-linked glycoproteins, we performed N-linked glycoproteomic profiling in ESCC tissues by two-dimensional gel electrophoresis (2-DE)-based and isobaric tags for relative and absolute quantification (iTRAQ) labeling-based mass spectrometry quantitation in parallel, followed by validation of candidate glycoprotein biomarkers by Western blot.
RESULTS 2-DE-based and iTRAQ labeling-based quantitation identified 24 and 402 differentially expressed N-linked glycoproteins, respectively, with 15 in common, demonstrating the outperformance of iTRAQ labeling-based quantitation over 2-DE and complementarity of these two approaches. Proteomaps showed the distinct compositions of functional categories between proteins and glycoproteins with differential expression associated with ESCC. Western blot analysis validated the up-regulation of total procathepsin D and high-mannose procathepsin D, and the down-regulation of total haptoglobin, high-mannose clusterin, and GlcNAc/sialic acid-containing fraction of 14-3-3ζ in ESCC tissues. The serum levels of glycosylated fractions of clusterin, prolinearginine-rich end leucine-rich repeat protein, and haptoglobin in patients with ESCC were remarkably higher than those in healthy controls.
CONCLUSION Our study provides insights into the aberrant N-linked glycoproteome associated with ESCC, which will be a valuable resource for future investigations.
Original language | English |
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Pages (from-to) | 3869-3885 |
Number of pages | 17 |
Journal | World Journal of Gastroenterology |
Volume | 28 |
Issue number | 29 |
DOIs | |
Publication status | Published - 7 Aug 2022 |
Bibliographical note
Funding Information:Supported by National Natural Science Foundation of China, No. 81072039 and No. 81872037.
Publisher Copyright:
© The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
Keywords
- 14-3-3ζ
- Cathepsin D
- Esophageal squamous cell carcinoma
- Haptoglobin
- Lectin
- N-linked glycoprotein
- Post-translational modification
ASJC Scopus subject areas
- Gastroenterology