Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1984 Jan 1;217(1):199–207. doi: 10.1042/bj2170199

Comparative study on glycosaminoglycans synthesized in peripheral and peritoneal polymorphonuclear leucocytes from guinea pigs.

Y Ohhashi, F Hasumi, Y Mori
PMCID: PMC1153197  PMID: 6696721

Abstract

Glycosaminoglycans synthesized in polymorphonuclear (PMN) leucocytes isolated from blood (peripheral PMN leucocytes) and in those induced intraperitoneally by the injection of caseinate (peritoneal PMN leucocytes) were compared. Both peripheral and peritoneal PMN leucocytes were incubated in medium containing [35S]sulphate and [3H]glucosamine. Each sample obtained after incubation was separated into cell, cell-surface and medium fractions by trypsin digestion and centrifugation. The glycosaminoglycans secreted from peripheral and peritoneal PMN leucocytes were decreased in size by alkali treatment, indicating that they existed in the form of proteoglycans. Descending paper chromatography of the unsaturated disaccharides obtained by the digestion of glycosaminoglycans with chondroitinase AC and chondroitinase ABC identified the labelled glycosaminoglycans of both the cell and the medium fractions in peripheral PMN leucocytes as 55-58% chondroitin 4-sulphate, 16-19% chondroitin 6-sulphate, 16-19% dermatan sulphate and 6-8% heparan sulphate. Oversulphated chondroitin sulphate and oversulphated dermatan sulphate were found only in the medium fraction. In peritoneal PMN leucocytes there is a difference in the composition of glycosaminoglycans between the cell and the medium fractions; the cell fraction was composed of 60% chondroitin 4-sulphate, 5.5% chondroitin 6-sulphate, 16.8% dermatan sulphate and 13.9% heparan sulphate, whereas the medium fraction consisted of 24.5% chondroitin 4-sulphate, 28.2% chondroitin 6-sulphate, 33.7% dermatan sulphate and 10% heparan sulphate. Oversulphated chondroitin sulphate and oversulphated dermatan sulphate were found in the cell, cell-surface and medium fractions. On the basis of enzymic assays with chondro-4-sulphatase and chondro-6-sulphatase, the positions of sulphation in the disulphated disaccharides were identified as 4- and 6-positions of N-acetylgalactosamine. Most of the 35S-labelled glycosaminoglycans synthesized in peripheral PMN leucocytes were retained within cells, whereas those in peritoneal PMN leucocytes were secreted into the culture medium. Moreover, the amount of glycosaminoglycans in peritoneal PMN leucocytes was significantly less than that in peripheral PMN leucocytes. Assay of lysosomal enzymes showed that these activities in peritoneal PMN leucocytes were 2-fold higher than those in peripheral PMN leucocytes.

Full text

PDF
207

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Avila J. L., Convit J. Studies on human polymorphonuclear leukocyte enzymes. I. Assay of acid hydrolases and other enzymes. Biochim Biophys Acta. 1973 Feb 15;293(2):397–408. doi: 10.1016/0005-2744(73)90347-1. [DOI] [PubMed] [Google Scholar]
  2. BITTER T., MUIR H. M. A modified uronic acid carbazole reaction. Anal Biochem. 1962 Oct;4:330–334. doi: 10.1016/0003-2697(62)90095-7. [DOI] [PubMed] [Google Scholar]
  3. Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968;97:77–89. [PubMed] [Google Scholar]
  4. HORN R. G., SPICER S. S. SULFATED MUCOPOLYSACCHARIDE AND BASIC PROTEIN IN CERTAIN GRANULES OF RABBIT LEUKOCYTES. Lab Invest. 1964 Jan;13:1–15. [PubMed] [Google Scholar]
  5. Hart G. W. Biosynthesis of glycosaminoglycans by thymic lymphocytes. Effects of mitogenic activation. Biochemistry. 1982 Nov 23;21(24):6088–6096. doi: 10.1021/bi00267a010. [DOI] [PubMed] [Google Scholar]
  6. Hasumi F., Mori Y. Characterization of glycosaminoglycans in peritoneal leukocytes from guinea pig. Biochem Med. 1980 Feb;23(1):6–16. doi: 10.1016/0006-2944(80)90049-6. [DOI] [PubMed] [Google Scholar]
  7. Kakinuma K. Improved method for obtaining polymorphonuclear leucocytes from guinea pigs. Jpn J Exp Med. 1968 Jun;38(3):165–169. [PubMed] [Google Scholar]
  8. Kawai Y., Seno N., Anno K. Chondroitin polysulfate of squid cartilage. J Biochem. 1966 Sep;60(3):317–321. doi: 10.1093/oxfordjournals.jbchem.a128438. [DOI] [PubMed] [Google Scholar]
  9. Kolset S. O., Kjellén L., Seljelid R., Lindahl U. Changes in glycosaminoglycan biosynthesis during differentiation in vitro of human monocytes. Biochem J. 1983 Mar 15;210(3):661–667. doi: 10.1042/bj2100661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Murata K. Acidic glycosaminoglycans in human platelets and leukocytes: the isolation and enzymatic characterization of chondroitin 4-sulfate. Clin Chim Acta. 1974 Dec 2;57(2):115–124. doi: 10.1016/0009-8981(74)90418-5. [DOI] [PubMed] [Google Scholar]
  11. Murata K., Ogura T., Okuyama T. The acidic glycosaminoglycans in leukocytes: an application of enzymatic methods. Connect Tissue Res. 1974;2(2):101–109. doi: 10.3109/03008207409152095. [DOI] [PubMed] [Google Scholar]
  12. Murata K. Structural differences of acidic glycosaminoglycans of leukocytes in leukemic states. Biochem Med. 1980 Jun;23(3):324–335. doi: 10.1016/0006-2944(80)90043-5. [DOI] [PubMed] [Google Scholar]
  13. Olsson I., Gardell S. Isolation and characterization of glycosaminoglycans from human leukocytes and platelets. Biochim Biophys Acta. 1967 Jul 25;141(2):348–357. doi: 10.1016/0304-4165(67)90109-2. [DOI] [PubMed] [Google Scholar]
  14. Olsson I. Intracellular distribution and sites of synthesis of glycosaminoglycans (mucopolysaccharides) in human leukocytes. Exp Cell Res. 1969 Mar;54(3):314–317. doi: 10.1016/0014-4827(69)90208-0. [DOI] [PubMed] [Google Scholar]
  15. Olsson I. Isolation of human leukocyte granules using colloidal silica-polysaccharide density gradients. Exp Cell Res. 1969 Mar;54(3):325–330. doi: 10.1016/0014-4827(69)90210-9. [DOI] [PubMed] [Google Scholar]
  16. Olsson I. The intracellular transport of glycosaminoglycans (mucopolysaccharides) in human leukocytes. Exp Cell Res. 1969 Mar;54(3):318–325. doi: 10.1016/0014-4827(69)90209-2. [DOI] [PubMed] [Google Scholar]
  17. Parmley R. T., Eguchi M., Spicer S. S. Ultrastructural cytochemistry of complex carbohydrates in leukocyte granules. J Histochem Cytochem. 1979 Aug;27(8):1167–1170. doi: 10.1177/27.8.90076. [DOI] [PubMed] [Google Scholar]
  18. Schuchman E. H., Desnick R. J. A new continuous, monodimensional electrophoretic system for the separation and quantitation of individual glycosaminoglycans. Anal Biochem. 1981 Nov 1;117(2):419–426. doi: 10.1016/0003-2697(81)90801-0. [DOI] [PubMed] [Google Scholar]
  19. Takasu Y., Hasumi F., Mori Y. Biosynthesis of glycosaminoglycans in peritoneal macrophages from the guinea pig. Biochim Biophys Acta. 1982 Jun 16;716(3):316–323. doi: 10.1016/0304-4165(82)90022-8. [DOI] [PubMed] [Google Scholar]
  20. Vannucchi S., Fibbi G., Cappelletti R., Del Rosso M., Chiarugi V. Glycosaminoglycan changes involved in polymorphonuclear leukocyte activation in vitro. J Cell Physiol. 1982 May;111(2):149–154. doi: 10.1002/jcp.1041110205. [DOI] [PubMed] [Google Scholar]
  21. Zeya H. I., Spitznagel J. K. Characterization of cationic protein-bearing granules of polymorphonuclear leukocytes. Lab Invest. 1971 Mar;24(3):229–236. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES