Preparation and Characterization of Heparan Sulfate-Derived Oligosaccharides to Investigate Protein-GAG Interaction and HS Biosynthesis Enzyme Activity

Methods Mol Biol. 2022:2303:121-137. doi: 10.1007/978-1-0716-1398-6_11.

Abstract

Heparan sulfate chains are complex and structurally diverse polysaccharides that interact with a large number of proteins, thereby regulating a vast array of biological functions. Understanding this activity requires obtaining oligosaccharides of defined structures. Here we describe methods for isolating, engineering, and characterizing heparan sulfate-derived oligosaccharides and approaches based on high-performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR), and bio-layer interferometry (BLI) to study their structures, modifications, and interactions.

Keywords: Chemoenzymatic approaches; Glycosaminoglycans; HPLC disaccharide analysis; Heparan sulfate; NMR-based structural studies; Protein–heparan sulfate interaction.

MeSH terms

  • Chromatography, High Pressure Liquid
  • Heparitin Sulfate
  • Oligosaccharides / chemistry*
  • Proteins

Substances

  • Oligosaccharides
  • Proteins
  • Heparitin Sulfate