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

de Luca, S.; Lohmander, Stefan LU ; Nilsson, Boel LU ; Hascall, V. C. and Caplan, A. I. (1980) In Journal of Biological Chemistry 255(13). p.6077-6083
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... (More)

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.

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organization
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publication status
published
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in
Journal of Biological Chemistry
volume
255
issue
13
pages
7 pages
publisher
American Society for Biochemistry and Molecular Biology
external identifiers
  • Scopus:0019305138
ISSN
0021-9258
language
English
LU publication?
yes
id
c8516e35-bb5e-4858-9792-048e1a96c58a
date added to LUP
2016-05-04 12:50:01
date last changed
2016-07-04 11:06:01
@misc{c8516e35-bb5e-4858-9792-048e1a96c58a,
  abstract     = {<p>The precursors, [ <sup>35</sup>S]sulfate and [2- <sup>3</sup>H]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 <sup>35</sup>S 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 [ <sup>3</sup>H]mannose originally in monomer, but no <sup>35</sup>S 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.</p>},
  author       = {de Luca, S. and Lohmander, Stefan and Nilsson, Boel and Hascall, V. C. and Caplan, A. I.},
  issn         = {0021-9258},
  language     = {eng},
  number       = {13},
  pages        = {6077--6083},
  publisher    = {ARRAY(0x5ae27a0)},
  series       = {Journal of Biological Chemistry},
  title        = {Proteoglycans from chick limb bud chondrocyte cultures. Keratan sulfate and oligosaccharides which contain mannose and sialic acid},
  volume       = {255},
  year         = {1980},
}