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. 1990 Dec 1;111(6):3177–3188. doi: 10.1083/jcb.111.6.3177

Interaction of the NG2 chondroitin sulfate proteoglycan with type VI collagen

PMCID: PMC2116373  PMID: 2269670

Abstract

The NG2 chondroitin sulfate proteoglycan is a membrane-associated molecule of approximately 500 kD with a core glycoprotein of 300 kD. Both the complete proteoglycan and a smaller quantity of the 300-kD core are immunoprecipitable with polyclonal and monoclonal antibodies against purified NG2. From some cell lines, the antibodies coprecipitate NG2 and type VI collagen, the latter appearing on SDS- PAGE as components of 140 and 250 kD under reducing conditions. The immunoprecipitation of type VI collagen does not seem to be due to recognition of the collagen by the antibodies, but rather to binding of the collagen to NG2. Studies on the NG2-type VI collagen complex suggest that binding between the two molecules is mediated by protein- protein interactions rather than by ionic interactions involving the glycosaminoglycans. Immunofluorescence double labeling in frozen sections of embryonic rat shows that NG2 and type VI collagen are colocalized in structures such as the intervertebral discs and arteries of the spinal column. In vitro the two molecules are highly colocalized on the surface of several cell lines. Treatment of these cells resulting in a change in the distribution of NG2 on the cell surface also causes a parallel change in type VI collagen distribution. Our results suggest that cell surface NG2 may mediate cellular interactions with the extracellular matrix by binding to type VI collagen.

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Selected References

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  1. Berman P., Gray P., Chen E., Keyser K., Ehrlich D., Karten H., LaCorbiere M., Esch F., Schubert D. Sequence analysis, cellular localization, and expression of a neuroretina adhesion and cell survival molecule. Cell. 1987 Oct 9;51(1):135–142. doi: 10.1016/0092-8674(87)90018-3. [DOI] [PubMed] [Google Scholar]
  2. Bonaldo P., Russo V., Bucciotti F., Doliana R., Colombatti A. Structural and functional features of the alpha 3 chain indicate a bridging role for chicken collagen VI in connective tissues. Biochemistry. 1990 Feb 6;29(5):1245–1254. doi: 10.1021/bi00457a021. [DOI] [PubMed] [Google Scholar]
  3. Bray D. F., Frank C. B., Bray R. C. Cytochemical evidence for a proteoglycan-associated filamentous network in ligament extracellular matrix. J Orthop Res. 1990 Jan;8(1):1–12. doi: 10.1002/jor.1100080102. [DOI] [PubMed] [Google Scholar]
  4. Bumol T. F., Reisfeld R. A. Unique glycoprotein-proteoglycan complex defined by monoclonal antibody on human melanoma cells. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1245–1249. doi: 10.1073/pnas.79.4.1245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cheifetz S., Andres J. L., Massagué J. The transforming growth factor-beta receptor type III is a membrane proteoglycan. Domain structure of the receptor. J Biol Chem. 1988 Nov 15;263(32):16984–16991. [PubMed] [Google Scholar]
  6. Chu M. L., Zhang R. Z., Pan T. C., Stokes D., Conway D., Kuo H. J., Glanville R., Mayer U., Mann K., Deutzmann R. Mosaic structure of globular domains in the human type VI collagen alpha 3 chain: similarity to von Willebrand factor, fibronectin, actin, salivary proteins and aprotinin type protease inhibitors. EMBO J. 1990 Feb;9(2):385–393. doi: 10.1002/j.1460-2075.1990.tb08122.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cole G. J., Burg M. Characterization of a heparan sulfate proteoglycan that copurifies with the neural cell adhesion molecule. Exp Cell Res. 1989 May;182(1):44–60. doi: 10.1016/0014-4827(89)90278-4. [DOI] [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. Dixit V. M., Grant G. A., Santoro S. A., Frazier W. A. Isolation and characterization of a heparin-binding domain from the amino terminus of platelet thrombospondin. J Biol Chem. 1984 Aug 25;259(16):10100–10105. [PubMed] [Google Scholar]
  10. Engvall E., Hessle H., Klier G. Molecular assembly, secretion, and matrix deposition of type VI collagen. J Cell Biol. 1986 Mar;102(3):703–710. doi: 10.1083/jcb.102.3.703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Frenette G. P., Ruddon R. W., Krzesicki R. F., Naser J. A., Peters B. P. Biosynthesis and deposition of a noncovalent laminin-heparan sulfate proteoglycan complex and other basal lamina components by a human malignant cell line. J Biol Chem. 1989 Feb 25;264(6):3078–3088. [PubMed] [Google Scholar]
  12. Gallagher J. T. The extended family of proteoglycans: social residents of the pericellular zone. Curr Opin Cell Biol. 1989 Dec;1(6):1201–1218. doi: 10.1016/s0955-0674(89)80072-9. [DOI] [PubMed] [Google Scholar]
  13. Gospodarowicz D., Ferrara N., Schweigerer L., Neufeld G. Structural characterization and biological functions of fibroblast growth factor. Endocr Rev. 1987 May;8(2):95–114. doi: 10.1210/edrv-8-2-95. [DOI] [PubMed] [Google Scholar]
  14. Hassell J. R., Kimura J. H., Hascall V. C. Proteoglycan core protein families. Annu Rev Biochem. 1986;55:539–567. doi: 10.1146/annurev.bi.55.070186.002543. [DOI] [PubMed] [Google Scholar]
  15. Hessle H., Engvall E. Type VI collagen. Studies on its localization, structure, and biosynthetic form with monoclonal antibodies. J Biol Chem. 1984 Mar 25;259(6):3955–3961. [PubMed] [Google Scholar]
  16. Hoffman S., Crossin K. L., Edelman G. M. Molecular forms, binding functions, and developmental expression patterns of cytotactin and cytotactin-binding proteoglycan, an interactive pair of extracellular matrix molecules. J Cell Biol. 1988 Feb;106(2):519–532. doi: 10.1083/jcb.106.2.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hubbard A. L., Cohn Z. A. The enzymatic iodination of the red cell membrane. J Cell Biol. 1972 Nov;55(2):390–405. doi: 10.1083/jcb.55.2.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hök M., Kjellén L., Johansson S. Cell-surface glycosaminoglycans. Annu Rev Biochem. 1984;53:847–869. doi: 10.1146/annurev.bi.53.070184.004215. [DOI] [PubMed] [Google Scholar]
  19. Kearney J. F., Radbruch A., Liesegang B., Rajewsky K. A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol. 1979 Oct;123(4):1548–1550. [PubMed] [Google Scholar]
  20. Keene D. R., Engvall E., Glanville R. W. Ultrastructure of type VI collagen in human skin and cartilage suggests an anchoring function for this filamentous network. J Cell Biol. 1988 Nov;107(5):1995–2006. doi: 10.1083/jcb.107.5.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kiss I., Deák F., Mestrić S., Delius H., Soos J., Dékány K., Argraves W. S., Sparks K. J., Goetinck P. F. Structure of the chicken link protein gene: exons correlate with the protein domains. Proc Natl Acad Sci U S A. 1987 Sep;84(18):6399–6403. doi: 10.1073/pnas.84.18.6399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  23. Lander A. D., Fujii D. K., Reichardt L. F. Laminin is associated with the "neurite outgrowth-promoting factors" found in conditioned media. Proc Natl Acad Sci U S A. 1985 Apr;82(7):2183–2187. doi: 10.1073/pnas.82.7.2183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Levine J. M., Beasley L., Stallcup W. B. The D1.1 antigen: a cell surface marker for germinal cells of the central nervous system. J Neurosci. 1984 Mar;4(3):820–831. doi: 10.1523/JNEUROSCI.04-03-00820.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Levine J. M., Card J. P. Light and electron microscopic localization of a cell surface antigen (NG2) in the rat cerebellum: association with smooth protoplasmic astrocytes. J Neurosci. 1987 Sep;7(9):2711–2720. doi: 10.1523/JNEUROSCI.07-09-02711.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Levine J. M., Stallcup W. B. Plasticity of developing cerebellar cells in vitro studied with antibodies against the NG2 antigen. J Neurosci. 1987 Sep;7(9):2721–2731. doi: 10.1523/JNEUROSCI.07-09-02721.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Martin G. R., Timpl R. Laminin and other basement membrane components. Annu Rev Cell Biol. 1987;3:57–85. doi: 10.1146/annurev.cb.03.110187.000421. [DOI] [PubMed] [Google Scholar]
  28. Neame P. J., Christner J. E., Baker J. R. Cartilage proteoglycan aggregates. The link protein and proteoglycan amino-terminal globular domains have similar structures. J Biol Chem. 1987 Dec 25;262(36):17768–17778. [PubMed] [Google Scholar]
  29. Nichols W. W., Murphy D. G., Cristofalo V. J., Toji L. H., Greene A. E., Dwight S. A. Characterization of a new human diploid cell strain, IMR-90. Science. 1977 Apr 1;196(4285):60–63. doi: 10.1126/science.841339. [DOI] [PubMed] [Google Scholar]
  30. Oldberg A., Ruoslahti E. Interactions between chondroitin sulfate proteoglycan, fibronectin, and collagen. J Biol Chem. 1982 May 10;257(9):4859–4863. [PubMed] [Google Scholar]
  31. Raff M. C., Williams B. P., Miller R. H. The in vitro differentiation of a bipotential glial progenitor cell. EMBO J. 1984 Aug;3(8):1857–1864. doi: 10.1002/j.1460-2075.1984.tb02059.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rapraeger A., Jalkanen M., Bernfield M. Cell surface proteoglycan associates with the cytoskeleton at the basolateral cell surface of mouse mammary epithelial cells. J Cell Biol. 1986 Dec;103(6 Pt 2):2683–2696. doi: 10.1083/jcb.103.6.2683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Ruoslahti E., Engvall E. Complexing of fibronectin glycosaminoglycans and collagen. Biochim Biophys Acta. 1980 Aug 13;631(2):350–358. doi: 10.1016/0304-4165(80)90308-6. [DOI] [PubMed] [Google Scholar]
  34. Ruoslahti E. Fibronectin and its receptors. Annu Rev Biochem. 1988;57:375–413. doi: 10.1146/annurev.bi.57.070188.002111. [DOI] [PubMed] [Google Scholar]
  35. Ruoslahti E. Proteoglycans in cell regulation. J Biol Chem. 1989 Aug 15;264(23):13369–13372. [PubMed] [Google Scholar]
  36. Ruoslahti E. Structure and biology of proteoglycans. Annu Rev Cell Biol. 1988;4:229–255. doi: 10.1146/annurev.cb.04.110188.001305. [DOI] [PubMed] [Google Scholar]
  37. Sakai L. Y., Keene D. R., Engvall E. Fibrillin, a new 350-kD glycoprotein, is a component of extracellular microfibrils. J Cell Biol. 1986 Dec;103(6 Pt 1):2499–2509. doi: 10.1083/jcb.103.6.2499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Saunders S., Bernfield M. Cell surface proteoglycan binds mouse mammary epithelial cells to fibronectin and behaves as a receptor for interstitial matrix. J Cell Biol. 1988 Feb;106(2):423–430. doi: 10.1083/jcb.106.2.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. 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]
  40. Schubert D., Heinemann S., Carlisle W., Tarikas H., Kimes B., Patrick J., Steinbach J. H., Culp W., Brandt B. L. Clonal cell lines from the rat central nervous system. Nature. 1974 May 17;249(454):224–227. doi: 10.1038/249224a0. [DOI] [PubMed] [Google Scholar]
  41. Scott J. E. Proteoglycan-fibrillar collagen interactions. Biochem J. 1988 Jun 1;252(2):313–323. doi: 10.1042/bj2520313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Segarini P. R., Seyedin S. M. The high molecular weight receptor to transforming growth factor-beta contains glycosaminoglycan chains. J Biol Chem. 1988 Jun 15;263(17):8366–8370. [PubMed] [Google Scholar]
  43. Smith G. N., Jr, Williams J. M., Brandt K. D. Interaction of proteoglycans with the pericellular (1 alpha, 2 alpha, 3 alpha) collagens of cartilage. J Biol Chem. 1985 Sep 5;260(19):10761–10767. [PubMed] [Google Scholar]
  44. Stallcup W. B., Beasley L. L., Levine J. M. Antibody against nerve growth factor-inducible large external (NILE) glycoprotein labels nerve fiber tracts in the developing rat nervous system. J Neurosci. 1985 Apr;5(4):1090–1101. doi: 10.1523/JNEUROSCI.05-04-01090.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Stallcup W. B., Beasley L. Bipotential glial precursor cells of the optic nerve express the NG2 proteoglycan. J Neurosci. 1987 Sep;7(9):2737–2744. doi: 10.1523/JNEUROSCI.07-09-02737.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Stallcup W. B., Beasley L., Levine J. Cell-surface molecules that characterize different stages in the development of cerebellar interneurons. Cold Spring Harb Symp Quant Biol. 1983;48(Pt 2):761–774. doi: 10.1101/sqb.1983.048.01.078. [DOI] [PubMed] [Google Scholar]
  47. Stallcup W. B. The NG2 antigen, a putative lineage marker: immunofluorescent localization in primary cultures of rat brain. Dev Biol. 1981 Apr 15;83(1):154–165. doi: 10.1016/s0012-1606(81)80018-8. [DOI] [PubMed] [Google Scholar]
  48. Sun X., Mosher D. F., Rapraeger A. Heparan sulfate-mediated binding of epithelial cell surface proteoglycan to thrombospondin. J Biol Chem. 1989 Feb 15;264(5):2885–2889. [PubMed] [Google Scholar]
  49. Suzuki S., Oldberg A., Hayman E. G., Pierschbacher M. D., Ruoslahti E. Complete amino acid sequence of human vitronectin deduced from cDNA. Similarity of cell attachment sites in vitronectin and fibronectin. EMBO J. 1985 Oct;4(10):2519–2524. doi: 10.1002/j.1460-2075.1985.tb03965.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Suzuki S., Pierschbacher M. D., Hayman E. G., Nguyen K., Ohgren Y., Ruoslahti E. Domain structure of vitronectin. Alignment of active sites. J Biol Chem. 1984 Dec 25;259(24):15307–15314. [PubMed] [Google Scholar]
  51. Vogel K. G., Keller E. J., Lenhoff R. J., Campbell K., Koob T. J. Proteoglycan synthesis by fibroblast cultures initiated from regions of adult bovine tendon subjected to different mechanical forces. Eur J Cell Biol. 1986 Jun;41(1):102–112. [PubMed] [Google Scholar]
  52. 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]

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