Domain structure elucidation of human decorin glycosaminoglycans

Biochem J. 2010 Oct 15;431(2):199-205. doi: 10.1042/BJ20100788.

Abstract

The structure of the GAG (glycosaminoglycan) chain of recombinantly expressed decorin proteoglycan was examined using a combination of intact-chain analysis and domain compositional analysis. The GAG had a number-average molecular mass of 22 kDa as determined by PAGE. NMR spectroscopic analysis using two-dimensional correlation spectroscopy indicated that the ratio of glucuronic acid to iduronic acid in decorin peptidoglycan was 5 to 1. GAG domains terminated with a specific disaccharide obtained by enzymatic degradation of decorin GAG with highly specific endolytic and exolytic lyases were analysed by PAGE and further depolymerized with the enzymes. The disaccharide compositional profiles of the resulting domains were obtained using LC with mass spectrometric and photometric detection and compared with that of the polysaccharide. The information obtained through the disaccharide compositional profiling was combined with the NMR and PAGE data to construct a map of the decorin GAG sequence motifs.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Chromatography, Liquid
  • Decorin
  • Disaccharides / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Extracellular Matrix Proteins / biosynthesis
  • Extracellular Matrix Proteins / chemistry*
  • Glycosaminoglycans / biosynthesis
  • Glycosaminoglycans / chemistry*
  • Humans
  • Mass Spectrometry
  • Molecular Sequence Data
  • Protein Structure, Tertiary
  • Proteoglycans / biosynthesis
  • Proteoglycans / chemistry*

Substances

  • DCN protein, human
  • Decorin
  • Disaccharides
  • Extracellular Matrix Proteins
  • Glycosaminoglycans
  • Proteoglycans