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Structure and synthesis of intracellular proteoglycan in HL-60 human leukemic promyelocytes

Stefan Lohmander, L.; Arnljots, Kristina LU and Yanagishita, Masaki (1990) In Journal of Biological Chemistry 265(10). p.5802-5808
Abstract

The structure, biosynthesis, and metabolism of proteoglycans in the HL-60 human promyelocytes were studied by metabolic labeling in culture with [35S]sulfate, [3H]glucosamine, [3H]serine, and [3H]leucine. These cells synthesize a single predominant species of intracellular proteoglycan with an approximate molecular weight of 100,000. The cells contain about 1 μg of proteoglycan/million cells. The proteoglycan is turned over within the cells in two apparent pools with half-lives of about 0.6 and 27 h, respectively. The fast pool represents secretion into medium in an apparently intact form, whereas the slow pool represents intracellular degradation to free chondroitin sulfate chains and smaller... (More)

The structure, biosynthesis, and metabolism of proteoglycans in the HL-60 human promyelocytes were studied by metabolic labeling in culture with [35S]sulfate, [3H]glucosamine, [3H]serine, and [3H]leucine. These cells synthesize a single predominant species of intracellular proteoglycan with an approximate molecular weight of 100,000. The cells contain about 1 μg of proteoglycan/million cells. The proteoglycan is turned over within the cells in two apparent pools with half-lives of about 0.6 and 27 h, respectively. The fast pool represents secretion into medium in an apparently intact form, whereas the slow pool represents intracellular degradation to free chondroitin sulfate chains and smaller fragments. The proteoglycan contains a protein core with an apparent Mr on gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of about 20,000-30,000. To the core protein are attached an average of six or seven chondroitin sulfate chains, each with an Mr of about 10,000. The chondroitin sulfate chains contain ∼85% 4-sulfated and ∼15% nonsulfated disaccharides. The chondroitin sulfate attachment region of the core protein is essentially resistant to trypsin and elastase, whereas the remainder of the protein core is readily degraded by proteases. The size of the chondroitin sulfate attachment region peptide generated by trypsin was estimated to be ∼5 kDa. Based on the molecular size, distribution of amino acids, protease susceptibility, and the extent of O-glycosylation, we propose that the intracellular proteoglycan characterized in this study is the translation product of a proteoglycan gene reported to be present in these cells

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
leukemic, proteoglycans
in
Journal of Biological Chemistry
volume
265
issue
10
pages
7 pages
publisher
American Society for Biochemistry and Molecular Biology
external identifiers
  • Scopus:0025265152
ISSN
0021-9258
language
English
LU publication?
yes
id
7ac8a86c-0215-4e45-b12d-7d4d5d09c4c4
date added to LUP
2016-05-04 18:15:06
date last changed
2016-11-08 15:05:27
@misc{7ac8a86c-0215-4e45-b12d-7d4d5d09c4c4,
  abstract     = {<p>The structure, biosynthesis, and metabolism of proteoglycans in the HL-60 human promyelocytes were studied by metabolic labeling in culture with [<sup>35</sup>S]sulfate, [<sup>3</sup>H]glucosamine, [<sup>3</sup>H]serine, and [<sup>3</sup>H]leucine. These cells synthesize a single predominant species of intracellular proteoglycan with an approximate molecular weight of 100,000. The cells contain about 1 μg of proteoglycan/million cells. The proteoglycan is turned over within the cells in two apparent pools with half-lives of about 0.6 and 27 h, respectively. The fast pool represents secretion into medium in an apparently intact form, whereas the slow pool represents intracellular degradation to free chondroitin sulfate chains and smaller fragments. The proteoglycan contains a protein core with an apparent M<sub>r</sub> on gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of about 20,000-30,000. To the core protein are attached an average of six or seven chondroitin sulfate chains, each with an Mr of about 10,000. The chondroitin sulfate chains contain ∼85% 4-sulfated and ∼15% nonsulfated disaccharides. The chondroitin sulfate attachment region of the core protein is essentially resistant to trypsin and elastase, whereas the remainder of the protein core is readily degraded by proteases. The size of the chondroitin sulfate attachment region peptide generated by trypsin was estimated to be ∼5 kDa. Based on the molecular size, distribution of amino acids, protease susceptibility, and the extent of O-glycosylation, we propose that the intracellular proteoglycan characterized in this study is the translation product of a proteoglycan gene reported to be present in these cells </p>},
  author       = {Stefan Lohmander, L. and Arnljots, Kristina and Yanagishita, Masaki},
  issn         = {0021-9258},
  keyword      = {leukemic,proteoglycans},
  language     = {eng},
  number       = {10},
  pages        = {5802--5808},
  publisher    = {ARRAY(0x9a7b158)},
  series       = {Journal of Biological Chemistry},
  title        = {Structure and synthesis of intracellular proteoglycan in HL-60 human leukemic promyelocytes},
  volume       = {265},
  year         = {1990},
}