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. 1989 Mar 1;108(3):1165–1173. doi: 10.1083/jcb.108.3.1165

Membrane-associated chondroitin sulfate proteoglycans of human lung fibroblasts

PMCID: PMC2115369  PMID: 2646307

Abstract

Cultured human fetal lung fibroblasts produce some chondroitin sulfate proteoglycans that are extracted as an aggregate in chaotropic buffers containing 4 M guanidinium chloride. The aggregated proteoglycans are excluded from Sepharose CL4B and 2B, but become included, eluting with a Kav value of 0.53 from Sepharose CL4B, when Triton X-100 is included in the buffer. Conversely, some of the detergent-extractable chondroitin sulfate proteoglycans can be incorporated into liposomes, suggesting the existence of a hydrophobic membrane-intercalated chondroitin sulfate proteoglycan fraction. Purified preparations of hydrophobic chondroitin sulfate proteoglycans contain two major core protein forms of 90 and 52 kD. A monoclonal antibody (F58-7D8) obtained from the fusion of myeloma cells with spleen cells of BALB/c mice that were immunized with hydrophobic proteoglycans recognized the 90- but not the 52-kD core protein. The epitope that is recognized by the antibody is exposed at the surface of cultured human lung fibroblasts and at the surface of several stromal cells in vivo, but also at the surface of Kupffer cells and of epidermal cells. The core proteins of these small membrane-associated chondroitin sulfate proteoglycans are probably distinct from those previously identified in human fibroblasts by biochemical, immunological, and molecular biological approaches.

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Selected References

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  1. Avnur Z., Geiger B. Immunocytochemical localization of native chondroitin-sulfate in tissues and cultured cells using specific monoclonal antibody. Cell. 1984 Oct;38(3):811–822. doi: 10.1016/0092-8674(84)90276-9. [DOI] [PubMed] [Google Scholar]
  2. Brennan M. J., Oldberg A., Pierschbacher M. D., Ruoslahti E. Chondroitin/dermatan sulfate proteoglycan in human fetal membranes. Demonstration of an antigenically similar proteoglycan in fibroblasts. J Biol Chem. 1984 Nov 25;259(22):13742–13750. [PubMed] [Google Scholar]
  3. Bumol T. F., Walker L. E., Reisfeld R. A. Biosynthetic studies of proteoglycans in human melanoma cells with a monoclonal antibody to a core glycoprotein of chondroitin sulfate proteoglycans. J Biol Chem. 1984 Oct 25;259(20):12733–12741. [PubMed] [Google Scholar]
  4. Carlson S. S., Wight T. N. Nerve terminal anchorage protein 1 (TAP-1) is a chondroitin sulfate proteoglycan: biochemical and electron microscopic characterization. J Cell Biol. 1987 Dec;105(6 Pt 2):3075–3086. doi: 10.1083/jcb.105.6.3075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carlstedt I., Cöster L., Malmström A. Isolation and characterization of dermatan sulphate and heparan sulphate proteoglycans from fibroblast culture. Biochem J. 1981 Jul 1;197(1):217–225. doi: 10.1042/bj1970217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cöster L., Carlstedt I., Malmström A. Isolation of 35S- and 3H-labelled proteoglycans from cultures of human embryonic skin fibroblasts. Biochem J. 1979 Dec 1;183(3):669–681. doi: 10.1042/bj1830669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cöster L., Carlstedt I., Malmström A., Särnstrand B. Biosynthesis and secretion of dermatan sulphate proteoglycans in cultures of human skin fibroblasts. Biochem J. 1984 Jun 1;220(2):575–582. doi: 10.1042/bj2200575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. David G., Van den Berghe H. Heparan sulfate-chondroitin sulfate hybrid proteoglycan of the cell surface and basement membrane of mouse mammary epithelial cells. J Biol Chem. 1985 Sep 15;260(20):11067–11074. [PubMed] [Google Scholar]
  9. Doege K., Sasaki M., Horigan E., Hassell J. R., Yamada Y. Complete primary structure of the rat cartilage proteoglycan core protein deduced from cDNA clones. J Biol Chem. 1987 Dec 25;262(36):17757–17767. [PubMed] [Google Scholar]
  10. Garrigues H. J., Lark M. W., Lara S., Hellström I., Hellström K. E., Wight T. N. The melanoma proteoglycan: restricted expression on microspikes, a specific microdomain of the cell surface. J Cell Biol. 1986 Nov;103(5):1699–1710. doi: 10.1083/jcb.103.5.1699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Giacoletto K. S., Sant A. J., Bono C., Gorka J., O'Sullivan D. M., Quaranta V., Schwartz B. D. The human invariant chain is the core protein of the human class II-associated proteoglycan. J Exp Med. 1986 Nov 1;164(5):1422–1439. doi: 10.1084/jem.164.5.1422. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Glössl J., Beck M., Kresse H. Biosynthesis of proteodermatan sulfate in cultured human fibroblasts. J Biol Chem. 1984 Nov 25;259(22):14144–14150. [PubMed] [Google Scholar]
  13. Glössl J., Schubert-Prinz R., Gregory J. D., Damle S. P., von Figura K., Kresse H. Receptor-mediated endocytosis of proteoglycans by human fibroblasts involves recognition of the protein core. Biochem J. 1983 Nov 1;215(2):295–301. doi: 10.1042/bj2150295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Harper J. R., Quaranta V., Reisfeld R. A. Ammonium chloride interferes with a distinct step in the biosynthesis and cell surface expression of human melanoma-type chondroitin sulfate proteoglycan. J Biol Chem. 1986 Mar 15;261(8):3600–3606. [PubMed] [Google Scholar]
  15. Hascall V. C., Heinegård D. Aggregation of cartilage proteoglycans. I. The role of hyaluronic acid. J Biol Chem. 1974 Jul 10;249(13):4232–4241. [PubMed] [Google Scholar]
  16. Hedman K., Christner J., Julkunen I., Vaheri A. Chondroitin sulfate at the plasma membranes of cultured fibroblasts. J Cell Biol. 1983 Oct;97(4):1288–1293. doi: 10.1083/jcb.97.4.1288. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hedman K., Kurkinen M., Alitalo K., Vaheri A., Johansson S., Hök M. Isolation of the pericellular matrix of human fibroblast cultures. J Cell Biol. 1979 Apr;81(1):83–91. doi: 10.1083/jcb.81.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Heinegård D., Björne-Persson A., Cöster L., Franzén A., Gardell S., Malmström A., Paulsson M., Sandfalk R., Vogel K. The core proteins of large and small interstitial proteoglycans from various connective tissues form distinct subgroups. Biochem J. 1985 Aug 15;230(1):181–194. doi: 10.1042/bj2300181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Heremans A., Cassiman J. J., Van den Berghe H., David G. Heparan sulfate proteoglycan from the extracellular matrix of human lung fibroblasts. Isolation, purification, and core protein characterization. J Biol Chem. 1988 Apr 5;263(10):4731–4739. [PubMed] [Google Scholar]
  20. Johansson S., Hedman K., Kjellén L., Christner J., Vaheri A., Hök M. Structure and interactions of proteoglycans in the extracellular matrix produced by cultured human fibroblasts. Biochem J. 1985 Nov 15;232(1):161–168. doi: 10.1042/bj2320161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kartner N., Alon N., Swift M., Buchwald M., Riordan J. R. Isolation of plasma membranes from human skin fibroblasts. J Membr Biol. 1977 Sep 14;36(2-3):191–211. doi: 10.1007/BF01868151. [DOI] [PubMed] [Google Scholar]
  22. Krusius T., Gehlsen K. R., Ruoslahti E. A fibroblast chondroitin sulfate proteoglycan core protein contains lectin-like and growth factor-like sequences. J Biol Chem. 1987 Sep 25;262(27):13120–13125. [PubMed] [Google Scholar]
  23. Krusius T., Ruoslahti E. Primary structure of an extracellular matrix proteoglycan core protein deduced from cloned cDNA. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7683–7687. doi: 10.1073/pnas.83.20.7683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lewandowska K., Choi H. U., Rosenberg L. C., Zardi L., Culp L. A. Fibronectin-mediated adhesion of fibroblasts: inhibition by dermatan sulfate proteoglycan and evidence for a cryptic glycosaminoglycan-binding domain. J Cell Biol. 1987 Sep;105(3):1443–1454. doi: 10.1083/jcb.105.3.1443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lories V., David G., Cassiman J. J., Van den Berghe H. Heparan sulfate proteoglycans of human lung fibroblasts. Occurrence of distinct membrane, matrix and secreted forms. Eur J Biochem. 1986 Jul 15;158(2):351–359. doi: 10.1111/j.1432-1033.1986.tb09758.x. [DOI] [PubMed] [Google Scholar]
  26. Lories V., De Boeck H., David G., Cassiman J. J., Van den Berghe H. Heparan sulfate proteoglycans of human lung fibroblasts. Structural heterogeneity of the core proteins of the hydrophobic cell-associated forms. J Biol Chem. 1987 Jan 15;262(2):854–859. [PubMed] [Google Scholar]
  27. McGuire E. A., Tollefsen D. M. Activation of heparin cofactor II by fibroblasts and vascular smooth muscle cells. J Biol Chem. 1987 Jan 5;262(1):169–175. [PubMed] [Google Scholar]
  28. Rauch U., Glössl J., Kresse H. Comparison of small proteoglycans from skin fibroblasts and vascular smooth-muscle cells. Biochem J. 1986 Sep 1;238(2):465–474. doi: 10.1042/bj2380465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rosamond S., Brown L., Gomez C., Braciale T. J., Schwartz B. D. Xyloside inhibits synthesis of the class II-associated chondroitin sulfate proteoglycan and antigen presentation events. J Immunol. 1987 Sep 15;139(6):1946–1951. [PubMed] [Google Scholar]
  30. Sant A. J., Cullen S. E., Schwartz B. D. Identification of a sulfate-bearing molecule associated with HLA class II antigens. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1534–1538. doi: 10.1073/pnas.81.5.1534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schafer I. A., Sitabkha L., Pandy M. Isolation and preliminary characterization of proteoglycan aggregates from cultured dermal fibroblasts. J Biol Chem. 1984 Feb 25;259(4):2321–2330. [PubMed] [Google Scholar]
  32. Schmidt G., Robenek H., Harrach B., Glössl J., Nolte V., Hörmann H., Richter H., Kresse H. Interaction of small dermatan sulfate proteoglycan from fibroblasts with fibronectin. J Cell Biol. 1987 Jun;104(6):1683–1691. doi: 10.1083/jcb.104.6.1683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Scott P. G., Winterbottom N., Dodd C. M., Edwards E., Pearson C. H. A role for disulphide bridges in the protein core in the interaction of proteodermatan sulphate and collagen. Biochem Biophys Res Commun. 1986 Aug 14;138(3):1348–1354. doi: 10.1016/s0006-291x(86)80431-4. [DOI] [PubMed] [Google Scholar]
  34. Van der Schueren B., Gasser D., Marynen P., Van Leuven F., David G., Cassiman J. J., Van den Berghe H. Polymorphous endocytotic organelles in the receptor-mediated endocytosis of gold-labelled alpha 2-macroglobulin complexes by human fibroblasts. J Cell Sci. 1985 Apr;75:411–421. doi: 10.1242/jcs.75.1.411. [DOI] [PubMed] [Google Scholar]
  35. Vogel K. G., Paulsson M., Heinegård D. Specific inhibition of type I and type II collagen fibrillogenesis by the small proteoglycan of tendon. Biochem J. 1984 Nov 1;223(3):587–597. doi: 10.1042/bj2230587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Vogel K. G., Peterson D. W. Extracellular, surface, and intracellular proteoglycans produced by human embryo lung fibroblasts in culture (IMR-90). J Biol Chem. 1981 Dec 25;256(24):13235–13242. [PubMed] [Google Scholar]
  37. Voss B., Glössl J., Cully Z., Kresse H. Immunocytochemical investigation on the distribution of small chondroitin sulfate-dermatan sulfate proteoglycan in the human. J Histochem Cytochem. 1986 Aug;34(8):1013–1019. doi: 10.1177/34.8.2426331. [DOI] [PubMed] [Google Scholar]
  38. Yanagishita M., Hascall V. C. Proteoglycans synthesized by rat ovarian granulosa cells in culture. Isolation, fractionation, and characterization of proteoglycans associated with the cell layer. J Biol Chem. 1984 Aug 25;259(16):10260–10269. [PubMed] [Google Scholar]
  39. de Boeck H., Lories V., David G., Cassiman J. J., van den Berghe H. Identification of a 64 kDa heparan sulphate proteoglycan core protein from human lung fibroblast plasma membranes with a monoclonal antibody. Biochem J. 1987 Nov 1;247(3):765–771. doi: 10.1042/bj2470765. [DOI] [PMC free article] [PubMed] [Google Scholar]

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