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PMID: 4786528 Published · ppublish English Journal Article

A novel assay for the biosynthesis of sulphated polysaccharide and its application to studies on the effects of somatomedin on cultured cells.

The Biochemical journal ·Vol. 136 ·No. 4 ·1973-12-00 ·Pages 1069-74

Wasteson A, Uthne K, Westermark B

Abstract

1. A simple method is described for the determination of small amounts of [(35)S]sulphated polysaccharide with 95-100% recovery in the range from 0.3 to 150mug of polysaccharide. 2. The method is based on precipitation with cetylpyridinium chloride of polysaccharide samples applied to filter paper. It is not significantly disturbed by the presence in the sample of a large excess of inorganic (35)SO(4) (2-). 3. Sulphated glucosamino- and galactosaminoglycans may be determined separately by treatment of the sample with chondroitinase ABC. 4. The method is applicable to the assay of [(35)S]sulphated polysaccharide biosynthesis in cell cultures. A stimulation of sulphate incorporation obeying a linear dose-response curve, was demonstrated in somatomedin-incubated fibroblast and glia cell cultures. 5. The described system provides a new assay method for somatomedin. 6. The stimulatory effect of somatomedin on the synthesis of [(35)S]sulphated polysaccharide appeared to be general, rather than specific, for a particular type of polysaccharide.

MeSH Terms
Cells, Cultured/drug effects Cetylpyridinium Chemical Precipitation Chondroitin Dose-Response Relationship, Drug Fibroblasts/metabolism Galactosamine/analysis Glucosamine/analysis Humans Lyases/pharmacology Methods Neuroglia/metabolism Polysaccharides/analysis,biosynthesis Somatomedins/pharmacology Sulfates/biosynthesis Sulfur Radioisotopes
Chemicals
Polysaccharides Somatomedins Sulfates Sulfur Radioisotopes Galactosamine Chondroitin Cetylpyridinium Lyases Glucosamine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Wasteson A
Uthne K
Westermark B
References (21)
21 references, click to expand
  1. Stimulation by insulin in vitro of incorporation of (35S)sulfate and (14C)leucine into protein-polysaccharide complexes, (3H)uridine into RNA, and (3H)thymidine into DNA of costal cartilage from hypophysectomized rats.
    Endocrinology. 1968 Mar;82(3):493-9 PMID: 4230520
  2. Purification and properties of bacterial chondroitinases and chondrosulfatases.
    J Biol Chem. 1968 Apr 10;243(7):1523-35 PMID: 5647268
  3. The defect in Hurler's and Hunter's syndromes: faulty degradation of mucopolysaccharide.
    Proc Natl Acad Sci U S A. 1968 Jun;60(2):699-706 PMID: 4236091
  4. Long term culture of normal and neoplastic human glia.
    Acta Pathol Microbiol Scand. 1968;74(4):465-86 PMID: 4313504
  5. Quantative determination of the sulphation factor activity in human serum.
    Acta Endocrinol (Copenh). 1970 Feb;63(2):338-50 PMID: 5468347
  6. Synthesis of acid mucopolysaccharides by glial tumor cells in tissue culture.
    Proc Natl Acad Sci U S A. 1970 Jun;66(2):495-9 PMID: 4247492
  7. Production of sulfated mucopolysaccharides by established cell lines of fibroblastic and nonfibroblastic origin.
    Biochim Biophys Acta. 1970 Oct 27;222(1):240-3 PMID: 4248801
  8. Production and secretion of chondroitin sulfates and dermatan sulfate by established mammalian cell lines.
    Biochem Biophys Res Commun. 1971 May 21;43(4):723-8 PMID: 4327487
  9. Properties of fractionated chondroitin sulphate from ox nasal septa.
    Biochem J. 1971 May;122(4):477-85 PMID: 5123881
  10. Somatomedin: proposed designation for sulphation factor.
    Nature. 1972 Jan 14;235(5333):107 PMID: 4550398
  11. Glycosaminoglycan synthesis in mouse mastocytoma.
    Biochem J. 1972 Feb;126(3):587-92 PMID: 4263037
  12. Regulation of skeletal growth by sulfation factor.
    Adv Intern Med. 1971;17:237-63 PMID: 4949905
  13. Mode of degradation of the chondroitin sulphate proteoglycan in rat costal cartilage.
    Biochem J. 1972 Dec;130(3):729-38 PMID: 4664930
  14. Human somatomedians. Purification and some studies on their biological actions.
    Acta Endocrinol Suppl (Copenh). 1973;175:1-35 PMID: 4542208
  15. On the metabolism of the sulfate group of chondroitinsulfuric acid.
    J Biol Chem. 1952 May;196(2):477-81 PMID: 12980990
  16. Crystalline papain. I. Preparation, specificity, and activation.
    J Biol Chem. 1954 Apr;207(2):515-31 PMID: 13163037
  17. Amino acid metabolism in mammalian cell cultures.
    Science. 1959 Aug 21;130(3373):432-7 PMID: 13675766
  18. Immunoassay of insulin with insulin-antibody precipitate.
    Biochem J. 1963 Jul;88:137-46 PMID: 13952078
  19. CLONAL GROWTH OF MAMMALIAN CELLS IN A CHEMICALLY DEFINED, SYNTHETIC MEDIUM.
    Proc Natl Acad Sci U S A. 1965 Feb;53:288-93 PMID: 14294058
  20. THE LIMITED IN VITRO LIFETIME OF HUMAN DIPLOID CELL STRAINS.
    Exp Cell Res. 1965 Mar;37:614-36 PMID: 14315085
  21. Aliphatic ammonium salts in the assay of acidic polysaccharides from tissues.
    Methods Biochem Anal. 1960;8:145-97 PMID: 14444237
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj1361069
Published
1973-12-00
Pages
1069-74
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1166058
Subset
IM
External Links
PubMed source DOI: 10.1042/bj1361069 PMC Full Text View journal details
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