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. 1982 Jun 1;203(3):691–698. doi: 10.1042/bj2030691

Immunodiffusion studies of the tryptic fragments of bovine nasal-cartilage proteoglycan monomer of high buoyant density

Harold D Keiser 1
PMCID: PMC1158285  PMID: 6180731

Abstract

Tryptic fragments of bovine nasal-cartilage proteoglycan, fractionated by dissociative density-gradient ultracentrifugation, were made to react by immunodiffusion against antiserum to a hyaluronidase-digest subfraction of cartilage proteoglycan monomer. This reaction produced two families of partly superimposed precipitin lines. One family was restricted to gradient fractions of medium or low buoyant density and included the immunoprecipitation reaction attributed to the hyaluronic acid-binding region of the cartilage proteoglycan monomer. The second family of precipitin lines was present alone in gradient fractions of high buoyant density. Immunodiffusion studies with antisera to relatively homogeneous keratan sulphate-rich and chondroitin sulphate-bearing fragment subfractions isolated from the gradient fraction of highest density indicated that both subfractions contained the antigenic determinants responsible for the second family of precipitin lines. Additional immunodiffusion studies, with the use of multispecific antisera to chondroitinase ABC digest and hyaluronidase digest of proteoglycan monomer, confirmed that the two subfractions shared antigenic determinants, and, in addition, indicated that these determinants were on one molecular species in the keratan sulphate-rich fragment subfraction and divided among at least three in the chondroitin sulphate-bearing fragment subfraction. Although an unprecedentedly large number of cartilage proteoglycan antigens could be recognized with the antisera employed in this cartilage proteoglycan antigens could be recognized with the antisera employed in this study, it was not possible to identify antigenic determinants unambiguously specific for the three structurally and functionally distinct regions of the cartilage proteoglycan monomer.

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Selected References

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

  1. Christner J. E., Caterson B., Baker J. R. Immunological determinants of proteoglycans. Antibodies against the unsaturated oligosaccharide products of chondroitinase ABC-digested cartilage proteoglycans. J Biol Chem. 1980 Aug 10;255(15):7102–7105. [PubMed] [Google Scholar]
  2. Hascall V. C. Interaction of cartilage proteoglycans with hyaluronic acid. J Supramol Struct. 1977;7(1):101–120. doi: 10.1002/jss.400070110. [DOI] [PubMed] [Google Scholar]
  3. Hascall V. C., Sajdera S. W. Proteinpolysaccharide complex from bovine nasal cartilage. The function of glycoprotein in the formation of aggregates. J Biol Chem. 1969 May 10;244(9):2384–2396. [PubMed] [Google Scholar]
  4. Heinegård D. K., Hascall V. C. Characteristics of the nonaggregating proteoglycans isolated from bovine nasal cartilage. J Biol Chem. 1979 Feb 10;254(3):927–934. [PubMed] [Google Scholar]
  5. Heinegård D., Hascall V. C. Aggregation of cartilage proteoglycans. 3. Characteristics of the proteins isolated from trypsin digests of aggregates. J Biol Chem. 1974 Jul 10;249(13):4250–4256. [PubMed] [Google Scholar]
  6. Heinegård D., Paulsson M., Inerot S., Carlström C. A novel low-molecular weight chondroitin sulphate proteoglycan isolated from cartilage. Biochem J. 1981 Aug 1;197(2):355–366. doi: 10.1042/bj1970355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Keiser H. D., Adlersberg J. B., Steinman H. M. Isolation and biochemical characterization of the tryptic fragments of bovine nasal-cartilage proteoglycan monomer of high buoyant density. Biochem J. 1982 Jun 1;203(3):683–689. doi: 10.1042/bj2030683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Keiser H. D., Hatcher V. B. A comparison of bovine nasal cartilage proteoglycan core protein produced by chondroitinase and hyaluronidase: the possible role of protease contaminants. Connect Tissue Res. 1977;5(3):147–155. doi: 10.3109/03008207709152265. [DOI] [PubMed] [Google Scholar]
  9. Keiser H., DeVito J. Immunochemical studies of fragments of bovine nasal cartilage proteoglycan subunit. Connect Tissue Res. 1974;2(4):273–282. doi: 10.3109/03008207409152256. [DOI] [PubMed] [Google Scholar]
  10. Keiser H. Immunological studies of the fragments of bovine cartilage proteoglycan produced by chondroitinase-trypsin digestion. Arch Biochem Biophys. 1975 Jun;168(2):622–629. doi: 10.1016/0003-9861(75)90294-5. [DOI] [PubMed] [Google Scholar]
  11. Keiser H., Shulman H. J., Sandson J. I. Immunochemistry of cartilage proteoglycan. Immunodiffusion and gel-electrophoretic studies. Biochem J. 1972 Jan;126(1):163–169. doi: 10.1042/bj1260163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. OUCHTERLONY O. Diffusion-in-gel methods for immunological analysis. II. Prog Allergy. 1962;6:30–154. doi: 10.1159/000313795. [DOI] [PubMed] [Google Scholar]
  13. Roughley P. J., Mason R. M. The electrophoretic heterogeneity of bovine nasal cartilage proteoglycans. Biochem J. 1976 Aug 1;157(2):357–367. doi: 10.1042/bj1570357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sparks K. J., Lever P. L., Goetinck P. F. Antibody binding of cartilage-specific proteoglycans. Arch Biochem Biophys. 1980 Feb;199(2):579–590. doi: 10.1016/0003-9861(80)90316-1. [DOI] [PubMed] [Google Scholar]
  15. Stanescu V., Maroteaux P., Sobczak E. Proteoglycan populations of baboon (Papio papio) articular cartilage. Biochem J. 1977 Apr 1;163(1):103–109. doi: 10.1042/bj1630103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Swann D. A., Powell S., Sotman S. The heterogeneity of cartilage proteoglycans. Isolation of different types of proteoglycans from bovine articular cartilage. J Biol Chem. 1979 Feb 10;254(3):945–954. [PubMed] [Google Scholar]
  17. Wieslander J., Heinegárd D. Immunochemical analysis of cartilage proteoglycans. Radioimmunoassay of the molecules and the substructures. Biochem J. 1980 Jun 1;187(3):687–694. doi: 10.1042/bj1870687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Wieslander J., Heinegård D. Immunochemical analysis of cartilage proteoglycans. Antigenic determinants of substructures. Biochem J. 1979 Apr 1;179(1):35–45. doi: 10.1042/bj1790035. [DOI] [PMC free article] [PubMed] [Google Scholar]

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