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Glycobiology. 2015 Feb;25(2):151-6. doi: 10.1093/glycob/cwu114. Epub 2014 Oct 21.

Profiling sulfation/epimerization pattern of full-length heparan sulfate by NMR following cell culture 13C-glucose metabolic labeling.

Author information

1
Institut de Biologie Structurale (IBS), Université Grenoble Alpes, Grenoble F-38027, France CNRS, Grenoble F-38027, France CEA, DSV, IBS, Grenoble F-38027, France.
2
Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, Sweden.
3
Institut de Biologie Structurale (IBS), Université Grenoble Alpes, Grenoble F-38027, France CNRS, Grenoble F-38027, France CEA, DSV, IBS, Grenoble F-38027, France pierre.gans@ibs.fr hugues.lortat-Jacob@ibs.fr.

Abstract

Through its ability to interact with proteins, heparan sulfate (HS) fulfills a large variety of functions. Protein binding depends on the level of HS sulfation and epimerization which are cell specific and dynamically regulated. Characterization of this molecule, however, has been restricted to oligosaccharide fragments available in large amount for structural investigation or to sulfate distribution through compositional analysis. Here we developed a (1)H-(13)C 2D NMR-based approach, directly performed on HS isolated from (13)C-labeled cells. By integrating the peak volumes measured at different chemical shifts, this non-destructive analysis allows us to determine both the sulfation and the iduronic/glucuronic profiles of the polysaccharide. Applied to wild-type and N-deacetylase/N-sulfotransferase-deficient fibroblasts as well as to epithelial cells differentiation, it also gives insights into the functional relationships existing between HS biosynthetic enzymes. This approach should be of significant interest to better understand HS changes that occur through physiologic regulations or during pathological development.

KEYWORDS:

13C-metabolic labeling; heparan sulfate; quantification of GlcA/IdoA epimeriation; sulfation pattern

PMID:
25335974
DOI:
10.1093/glycob/cwu114
[Indexed for MEDLINE]

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