Proteoglycans from chick limb bud chondrocyte cultures. Keratan sulfate and oligosaccharides which contain mannose and sialic acid

Research output: Contribution to journalArticle

Abstract

The precursors, [ 35S]sulfate and [2- 3H]mannose, were used to study the biosynthesis of keratan sulfate and other oligosaccharides on proteoglycans isolated from Day 8 cultures of chick limb bud chondrocytes. After alkaline borohydride treatment, three fractions with sialic acid were separated by molecular sieve chromatography. The first contained keratan sulfate which was purified by digestion with chondroitinase to remove chondroitin sulfate, followed by molecular sieve and ion exchange chromatography. The purified keratan sulfate contained about 8% of the 35S activity originally in monomer. The chains had an average length of about 40 monosaccharides and contained only trace amounts of mannose (less than 1 residue/three to four chains). The second fraction contained the majority of the [ 3H]mannose originally in monomer, but no 35S activity. This fraction appears to contain oligosaccharide-peptides of the asparagine-N-glycosylamine type because there were no reduced sugars present and the alkaline borohydride treatment extensively degraded the core protein. The composition of the oligosaccharides, with high proportions of mannose, N-acetylglucosamine, galactose, and sialic acid, was consistent with this suggestion. The third fraction consisted of a series of oligosaccharides with sizes between three to six saccharides. They contained N-acetylgalactosaminitol, indicating that they were attached to the core protein by O-glycoside bonds between N-acetylgalactosamine and hydroxyl groups on serine and threonine. Thus, proteoglycans contain two classes of oligosaccharides, a mannose-rich class characteristic of glycoproteins and an O-glycoside class characteristic of mucins, in addition to the chondroitin sulfate and keratan sulfate chains.

Details

Authors
Organisations
External organisations
  • National Institutes of Health, United States
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Orthopedics
Original languageEnglish
Pages (from-to)6077-6083
Number of pages7
JournalJournal of Biological Chemistry
Volume255
Issue number13
Publication statusPublished - 1980
Publication categoryResearch
Peer-reviewedYes