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Persson A, Nikpour M, Vorontsov E, et al. Domain Mapping of Chondroitin/Dermatan Sulfate Glycosaminoglycans Enables Structural Characterization of Proteoglycans. Mol Cell Proteomics. 2021;20:100074doi: 10.1016/j.mcpro.2021.100074.
Persson, A., Nikpour, M., Vorontsov, E., Nilsson, J., & Larson, G. (2021). Domain Mapping of Chondroitin/Dermatan Sulfate Glycosaminoglycans Enables Structural Characterization of Proteoglycans. Molecular & cellular proteomics : MCP, 20100074. https://doi.org/10.1016/j.mcpro.2021.100074
Persson, Andrea, et al. "Domain Mapping of Chondroitin/Dermatan Sulfate Glycosaminoglycans Enables Structural Characterization of Proteoglycans." Molecular & cellular proteomics : MCP vol. 20 (2021): 100074. doi: https://doi.org/10.1016/j.mcpro.2021.100074
Persson A, Nikpour M, Vorontsov E, Nilsson J, Larson G. Domain Mapping of Chondroitin/Dermatan Sulfate Glycosaminoglycans Enables Structural Characterization of Proteoglycans. Mol Cell Proteomics. 2021;20:100074. doi: 10.1016/j.mcpro.2021.100074. Epub 2021 Mar 20. PMID: 33757834; PMCID: PMC8724862.
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Mol Cell Proteomics. 2021;20:100074. doi: 10.1016/j.mcpro.2021.100074. Epub 2021 Mar 20.

Domain Mapping of Chondroitin/Dermatan Sulfate Glycosaminoglycans Enables Structural Characterization of Proteoglycans.

Molecular & cellular proteomics : MCP

Andrea Persson, Mahnaz Nikpour, Egor Vorontsov, Jonas Nilsson, Göran Larson

Affiliations

  1. Department of Laboratory Medicine, Sahlgrenska Academy at the University of Gothenburg, Sweden. Electronic address: [email protected].
  2. Department of Laboratory Medicine, Sahlgrenska Academy at the University of Gothenburg, Sweden.
  3. Proteomics Core Facility, Sahlgrenska Academy at the University of Gothenburg, Sweden.
  4. Department of Laboratory Medicine, Sahlgrenska Academy at the University of Gothenburg, Sweden; Proteomics Core Facility, Sahlgrenska Academy at the University of Gothenburg, Sweden; Laboratory of Clinical Chemistry, Sahlgrenska University Hospital, Västra Götaland Region, Sweden.
  5. Department of Laboratory Medicine, Sahlgrenska Academy at the University of Gothenburg, Sweden; Laboratory of Clinical Chemistry, Sahlgrenska University Hospital, Västra Götaland Region, Sweden. Electronic address: [email protected].

PMID: 33757834 PMCID: PMC8724862 DOI: 10.1016/j.mcpro.2021.100074

Abstract

Of all posttranslational modifications known, glycosaminoglycans (GAGs) remain one of the most challenging to study, and despite the recent years of advancement in MS technologies and bioinformatics, detailed knowledge about the complete structures of GAGs as part of proteoglycans (PGs) is limited. To address this issue, we have developed a protocol to study PG-derived GAGs. Chondroitin/dermatan sulfate conjugates from the rat insulinoma cell line, INS-1832/13, known to produce primarily the PG chromogranin-A, were enriched by anion-exchange chromatography after pronase digestion. Following benzonase and hyaluronidase digestions, included in the sample preparation due to the apparent interference from oligonucleotides and hyaluronic acid in the analysis, the GAGs were orthogonally depolymerized and analyzed using nano-flow reversed-phase LC-MS/MS in negative mode. To facilitate the data interpretation, we applied an automated LC-MS peak detection and intensity measurement via the Proteome Discoverer software. This approach effectively provided a detailed structural description of the nonreducing end, internal, and linkage region domains of the CS/DS of chromogranin-A. The copolymeric CS/DS GAGs constituted primarily consecutive glucuronic-acid-containing disaccharide units, or CS motifs, of which the N-acetylgalactosamine residues were 4-O-sulfated, interspersed by single iduronic-acid-containing disaccharide units. Our data suggest a certain heterogeneity of the GAGs due to the identification of not only CS/DS GAGs but also of GAGs entirely of CS character. The presented protocol allows for the detailed characterization of PG-derived GAGs, which may greatly increase the knowledge about GAG structures in general and eventually lead to better understanding of how GAG structures are related to biological functions.

Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.

Keywords: LC-MS/MS; chondroitin/dermatan sulfate (CS/DS); glycan; glycomics; higher-energy collision dissociation (HCD); mass spectrometry (MS)

Conflict of interest statement

Conflict of interest The authors declare no competing interests.

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