Skip to main content

Advertisement

SpringerLink
Log in

The isolation of two types of collagen from embryonic bovine epiphyseal cartilage

  • Original Papers
  • Published:
Calcified Tissue Research Aims and scope Submit manuscript

Abstract

The nature of the collagen in embryonic and postnatal bovine epiphyseal cartilage was studied by amino acid analysis, extraction in denaturants, and extraction in denaturants after prior digestion of the cartilage with pepsin. Components of both Type I collagen and Type II collagen were isolated by molecular sieve filtration from the gelatin extracted from 4–5 month old embryonic bovine epiphyseal cartilage, and characterized by their behavior during ion exchange chromatography, amino acid composition and their content of hydroxylysine and glycosydically substituted hydroxylysine residues. Type I collagen accounted for about one-third of the total collagen content of epiphyseal cartilage. It was preferentially extracted in denaturant salts, whilst Type II collagen, essentially insoluble in denaturants, could be extracted from the cartilage only after prior digestion of the cartilage with pepsin. The marked preponderance of α components compared with higher molecular weight polymers in the extracted Type I gelatin suggested a paucity of intramolecular crosslinkages, and the lability of the intermolecular crosslinks to denaturants at neutral pH, suggests that the intermolecular, aldehyde-derived crosslinks of Type, I collagen in cartilage are similar to those in the Type I collagen of bone rather than those present in the Type I collagen of skin and other non-mineralized connective tissues. The marked differences in the solubility properties of Type I and Type II collagens of cartilage suggests that the stability and possibly the chemical nature of the intermolecular crosslinkages in Type II collagen, as well as the number of intramolecular crosslinkages, differs in the two types of collagen present in cartilage.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Aronson, R. B., Sinex, F. M., Franzblau, C., van Slyke, D.: The oxidation of protein-bound hydroxylysine by periodate. J. biol. Chem.242, 809–812 (1967).

    PubMed  CAS  Google Scholar 

  2. Bailey, A. J.: Comparative studies on the nature of the crosslinks stabilizing the collagen fibres of invertebrates, cyclostomes and elasmobranchs. FEBS Letters18, 154–158 (1971).

    Article  PubMed  CAS  Google Scholar 

  3. Bailey, A. J., Peach, C. M.: The chemistry of the collagen cross-links. Biochem. J.121, 257–259 (1971).

    PubMed  CAS  Google Scholar 

  4. Bailey, A. J., Peach, C. M., Fowler, L. J.: The biosynthesis of intermolecular crosslinks in collagen. In: The chemistry and molecular biology of the intercellular matrix, (Balazs, E. A., ed.), vol. 1, p. 385–404. London and New York: Academic Press 1970.

    Google Scholar 

  5. Bailey, A. J., Shimokomaki, M. S.: Age related changes in the reducible crosslinks of collagen. FEBS Letters16, 86–88 (1971).

    Article  PubMed  CAS  Google Scholar 

  6. Bonar, L. E., Glimcher, M. J.: A comparison of the organization and interaction properties of the collagen in bone and tendon as measured by wide and low-angle x-ray diffraction. J. Bone Jt Surg. A54, 1129 (1972).

    Google Scholar 

  7. Davis, N. R., Bailey, A. J.: Chemical synthesis of the reduced form of an intermolecular crosslink of collagen: A re-evaluation of the structure of syndesine. Biochem. biophys. Res. Commun.45, 1416–1422 (1971).

    Article  PubMed  CAS  Google Scholar 

  8. De Luque, O., Mechanic, G., Tanzer, M. L.: Isolation of peptides containing the crosslink, hydroxylysinonorleucine, from reconstituted collagen fibrils. Biochemistry9, 4987–4998 (1970).

    Article  PubMed  Google Scholar 

  9. Deshmukh, K., Mimni, M. E.: Isolation and characterization of cyanogen bromide peptides from the collagen of bovine articular cartilage. Biochem. J.133, 615–622 (1973).

    PubMed  CAS  Google Scholar 

  10. Eyre, D. R.: Reducible groups and the aggregation of extracted bone gelatin. J. Bone Jt. Surg. A54, 1129 (1972).

    Google Scholar 

  11. Eyre, D. R., Glimcher, M. J.: Evidence for a previously undetected sequence at the carboxyterminus of the α1 chain of chicken bone collagen. Biochem. biophys Res. Commun.48, 720–728 (1972).

    Article  PubMed  CAS  Google Scholar 

  12. Eyre, D. R., Glimcher, M. J.: Reducible crosslinks in the hydroxylysine deficient collagens of a heritable disorder of connective tissue. Proc. nat. Acad. Sci. (Wash.)69, 2594–2598 (1972).

    Article  CAS  Google Scholar 

  13. Eyre, D. R., Glimcher, M. J.: The distribution of crosslinking aldehydes in α1 and α2 chains of chicken bone collagen. Biochim. biophys. Acta (Amst.)278, 206–210 (1972).

    CAS  Google Scholar 

  14. Eyre, D. R., Glimcher, M. J.: Crosslinking in bone collagen—the sites of intermolecular and intramolecular aldehyde-mediated boods. J. Bone Jt Surg. A55, 651 (1973).

    Google Scholar 

  15. Eyre, D. R., Glimcher, M. J.: Evidence for intramolecular crosslinks in chicken bone collagen: the isolation of peptides containing allysine aldol. Biochim. biophys. Acta (Amst.)295, 301–307 (1973).

    CAS  Google Scholar 

  16. Eyre, D. R., Glimcher, M. J.: Collagen crosslinks: stability of aldiminiebonds in bone collagen. Proc. III Europ. Symp. on Connective tissue Research. Turku, Finland (in press)

  17. Eyre, D. R., Glimcher, M. J.: Dissolution of bovine and chicken bone collagens in concentrated formic acid. Calcif. Tiss. Res.15, 125–132 (1974).

    Article  CAS  Google Scholar 

  18. Eyre, D. R., Glimcher, M. J.: The lability of aldimine crosslinks on dissolution of chicken bone collagen by protein denaturants. Proc. 10th Europ. Symp. on Calc. Tiss., Hamburg, Germany (in press)

  19. Fell, H. B.: The histogenesis of cartilage and bone in the long bones of the embryonic fowl. J. Morph. Physiol.40, 417–459 (1925).

    Article  Google Scholar 

  20. Furthmayr, H., Timpl, R.: Characterization of collagen peptides by sodium dodecylsulfate polyacrylamide electrophoresis. Anal. Biochem.41, 510–516 (1971).

    Article  PubMed  CAS  Google Scholar 

  21. Gallop, P. M., Bulmenfeld, O. O., Seifter, S.: Structure and metabolism of connective tissue proteins. Ann. Rev. Biochem.41, 617–672 (1972)

    Article  PubMed  CAS  Google Scholar 

  22. Glimcher, M. J., Katz, E. R.: The organization of collagen in bone: The role of the noncovalent bonds in the relative insolubility of bone collagen. J. Ultrastruct Res.12, 705, 729 (1965).

    Google Scholar 

  23. Glimcher, M. J., Krane, S. M.: The organization and structure of bone and the mechanism of calcification. In: Treatise on collagen, vol. IIB, Biology of collagen, (Ramachandran, G. N., Gould, B. S., eds.) p. 68–251, London: Academic Press 1968.

    Google Scholar 

  24. Glimcher, M. J., Seyer, J. M., Brickley, D. M.: The solubilization of collagen and protein-polysaccharides from the developing cartilage of lathyritic chicks. Biochem. J.115, 923–926 (1969).

    PubMed  CAS  Google Scholar 

  25. Grant, R. A.: Estimation of hydroxyproline by the autoanalyzer. J. clin. Path.17, 685–689 (1969).

    Google Scholar 

  26. Linsenmayer, T. F., Toole, B. P., Trelstad, R. L.: Temporal and spatial transitions in collagen during embryonic chick limb development. Develop. Biol.35, 232–239 (1973).

    Article  PubMed  CAS  Google Scholar 

  27. Mechanic, G., Gallop, P. M., Tanzer, M. L.: The nature of crosslinking in collagens from mineralized tissues. Biochem. biophys. Res. Commun.45, 644–653 (1971).

    Article  PubMed  CAS  Google Scholar 

  28. Miller, E. J.: Isolation and characterization of a collagen from chick cartilage containing three identical alpha chains. Biochemistry10, 1652–1658 (1971).

    Article  PubMed  CAS  Google Scholar 

  29. Miller, E. J.: Isolation and characterization of the cyanogen bromide peptides from the α1(II) chain of chick cartilage collagen Biochemistry10, 3030–3035 (1971).

    Article  PubMed  CAS  Google Scholar 

  30. Miller, E. J.: Structural studies on cartilage collagen employing limited cleavage and solubilization with pepsin. Biochemistry11 4903–4909 (1972).

    Article  PubMed  CAS  Google Scholar 

  31. Miller, E. J.: A review of biochemical studies on the genetically distinct collagens of the skeletal system. Clin. Orthop.92, 260–280 (1973).

    Article  PubMed  Google Scholar 

  32. Miller, E. J., Epstein, E. H., Piez, K. A.: Identification of three genetically distinct collagens by cyanogen bromide cleavage of insoluble human skin and cartilage collagen. Biochem. biophys. Res. Commun.42, 1024–1029 (1971).

    Article  PubMed  CAS  Google Scholar 

  33. Miller, E. J., Lunde, L. G.: Isolation and Characterization of the Cyanogen Bromide Peptides from the α1(II) Chain of Bovine and Human Cartilage Collagen Biochemistry12, 3153–3159 (1973).

    Article  PubMed  CAS  Google Scholar 

  34. Miller, E. J., Matukas, V. J.: Chick cartilage collagen: A new type of α1 chain not present in bone or skin of the species. Proc. nat. Acad. Sci. (Wash.)64, 1264–1268 (1969).

    Article  CAS  Google Scholar 

  35. Miller, E. J., van der Korst, J. K., Sokoloff, L.: Collagen of human articular and costal cartilage. Arthr. and Rheum.12, 21–29 (1969).

    CAS  Google Scholar 

  36. Nimni, M., Deshmukh, K.: Differences in collagen metabolism between normal and osteoarthritic human articular cartilage. Science181, 751–752 (1973).

    Article  PubMed  CAS  Google Scholar 

  37. Pinnell, S. R., Fox, R., Krane, S. M.: Human collagens: Differences in glycosylated hydroxylysines in skin and bone. Biochim. biophys. Acta (Amst.)229, 119–122 (1971).

    CAS  Google Scholar 

  38. Pinnell, S. R., Krane, S. M., Kenzora, J. E., Glimcher, M. J.: A new heritable disorder of connective tissue. Hydroxylysine-deficient collagen disease. New Engl. J. Med.286, 1013–1020 (1972).

    Article  PubMed  CAS  Google Scholar 

  39. Rosenberg, L.: Chemical basis for the histological use of Saffranin-O in the study of articular cartilage. J. Bone Jt Surg. A53, 69–82 (1971).

    CAS  Google Scholar 

  40. Seyer, J. M., Brickley, D. M., Glimcher, M. J.: Investigation of two types of collagen in articular and epiphyseal cartilage. J. Bone Jt Surg. A54, 1130 (1972).

    Google Scholar 

  41. Stark, M., Rauterberg, J., Kuhn, K.: Evidence of a non-helical region at the carboxyl terminus of the collagen molecule. FEBS Letters13, 101–105 (1971).

    Article  PubMed  CAS  Google Scholar 

  42. Strawich, E., Nimni, M. E.: Properties of a collagen molecule containing three identical alpha components extracted from bovine articular cartilage. Biochemistry10, 3905–3911 (1971)

    Article  PubMed  CAS  Google Scholar 

  43. Tanzer, M. L., Mechanic, G., Gallop, P. M.: Isolation of hydroxylysinonorleucine and its lactone from reconstituted collagen fibrils. Biochim. biophys. Acta (Amst.)207, 548–552 (1970).

    CAS  Google Scholar 

  44. Toole, B. P., Kang, A. H., Trelstad, R. L., Gross, J.: Collagen heterogeneity within different growth regions of long bones of rachitic and non-rachitic chicks. Biochem. J.127, 715–720 (1972).

    PubMed  CAS  Google Scholar 

  45. Traub, W., Piez, K. A.: The chemistry and structure of collagen. Advanc. Protein Chem.25, 243–352 (1971).

    Article  CAS  Google Scholar 

  46. Trelstad, R. L., Kang, A. H., Igarashi, S., Gross, J.: Isolation of two distinct collagens from chick cartilage. Biochemistry9, 4993–4998 (1970).

    Article  PubMed  CAS  Google Scholar 

  47. Trelstad, R. L., Kang, A. H., Toole, B. P., Gross, J.: Collagen heterogeneity: High resolution separation of native [α1(I)]2α2 and [α1(II)]3 and their component chains J. biol. Chem.247, 6469 (1972).

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Orthopaedic Surgery, Harvard Medical School, The Children’s Hospital Medical Center, 300 Longwood Avenue, 02115, Boston, Massachusetts, USA

    J. M. Seyer M.D., D. M. Brickley & M. J. Glimcher

Authors
  1. J. M. Seyer M.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D. M. Brickley
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. J. Glimcher
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Seyer, J.M., Brickley, D.M. & Glimcher, M.J. The isolation of two types of collagen from embryonic bovine epiphyseal cartilage. Calc. Tis Res. 17, 25–41 (1974). https://doi.org/10.1007/BF02547212

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02547212

Key words

Search

Navigation