Calcified Tissue International

, Volume 33, Issue 1, pp 255–260

Uptake of collagenolytic enzymes by bone cells during isolation from embryonic rat calvaria

  • Jaro Sodek
  • Johan N. M. Heersche
Article

Summary

Collagenolytic activity extracted from bone cells, isolated from calvaria with a proteolytic enzyme mixture, was analyzed using14C-labeled collagen and a combination of gel electrophoresis and fluorography. The pattern of collagen degradation products obtained did not include 3/4-fragments, typical of vertebrate collagenases, but was essentially identical to the patterns generated by very dilute samples of the proteolytic enzyme mixture used to isolate the bone cells, and purified collagenase fromClostridium histolyticum (E.C. 3.4.24.3.). These and other results strongly suggest that the previously reported collagenolytic activity of bone cells [1] is exogenous in origin.

Key words

Bacterial collagenase Vertebrate collagenase Sodium dodecyl sulfate polyacrylamide gel electrophoresis Fluorography 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Puzas, J. E., Brand, J. S.: Collagenolytic activity from isolated bone cells, Biochem. Biophys. Acta429:946–974, 1976Google Scholar
  2. 2.
    Peck, W. A., Birge, S. J., Fedak, S. A.: Bone cells: biochemical and biological studies after enzymatic isolation, Science146:1476–1477, 1964PubMedGoogle Scholar
  3. 3.
    Dziak, R., Brand, J. S.: Calcium transport in isolated bone cells, J. Cell. Physiol.84:75–83, 1974CrossRefPubMedGoogle Scholar
  4. 4.
    Binderman, I., Duksin, D., Harell, A., Sachs, L., Katzir, E.: Formation of bone tissue in culture from isolated bone cells, J. Cell Biol.61:427–439, 1974CrossRefPubMedGoogle Scholar
  5. 5.
    Wong, G. L., Cohn, D. V.: Target cells in bone for parathormone and calcitonin are different: enrichment for each cell type by sequential digestion of mouse calvaria and selective adhesion to polymeric surfaces, Proc. Natl. Acad. Sci. U.S.A.72:3167–3171, 1975PubMedGoogle Scholar
  6. 6.
    Rao, L. G., Ng, B., Brunette, D. M., Heersche, J. N. M.: Parathyroid hormone- and prostaglandin E1-response in a selected population of bone cells after repeated subculture and storage at −80°C, Endocrinology100:1233–1241, 1977PubMedGoogle Scholar
  7. 7.
    Luben, R. A., Wong, G. L., Cohn, D. V.: Biochemical characterization with parathormone and calcitonin of isolated bone cells: provisional identification of osteoclasts and osteoblasts, Endocrinology99:526–534, 1976PubMedCrossRefGoogle Scholar
  8. 8.
    Puzas, J. E., Brand, J. S.: Parathyroid hormone stimulation of collagenase secretion by isolated bone cells, Endocrinology104:559–562, 1979PubMedGoogle Scholar
  9. 9.
    Rice, R. H., Means, G. E.: Radioactive labelling of proteinsin vitro, J. Biol. Chem.246:831–832, 1971PubMedGoogle Scholar
  10. 10.
    Pettigrew, D. W., Sodek, J., Wang, H.-M., Brunette, D. M.: Inhibitors of collagenolytic enzymes synthesised by fibroblasts and epithelial cells from porcine and macaque periodontal tissues, Arch. Oral Biol.25:269–274, 1980CrossRefPubMedGoogle Scholar
  11. 11.
    Laemmli, U. K.: Cleavage of structural proteins during the assembly of the head of the bacteriophage T4, Nature227:680–685, 1970CrossRefPubMedGoogle Scholar
  12. 12.
    Bonner, W. M., Laskey, R. A.: A film detection method for tritium labelled proteins and nucleic acids in polyacrylamide gels, Eur. J. Biochem.56:335–341, 1974Google Scholar
  13. 13.
    Pettigrew, D. W., Ho, G.-H., Sodek, J., Brunette, D. M., Wang, H.-M.: The effect of oxygen tension and indomethacin on production of collagenase and neutral proteinase enzymes and their latent forms by porcine gingival explants in culture, Arch. Oral Biol.23:767–777, 1978CrossRefPubMedGoogle Scholar
  14. 14.
    Peterkovsky, B., Diegelmann, R.: Use of a mixture of proteinase-free collagenases for the specific assay of radio-active collagen in the presence of other proteins, Biochemistry10:988–994, 1971CrossRefGoogle Scholar
  15. 15.
    Guzelian, P. S., Diegelmann, R. F.: Retention of clostridial collagenase by primary cultures of parenchymal cells prepared from adult rat liver, Life Sci.24:513–518, 1979CrossRefPubMedGoogle Scholar
  16. 16.
    Capuzzi, D. M., Sparks, C. E., DeHoff, J. L.: Effect of residual enzymes on degradation of radioiodinated VLDL by collagenase-dispersed hepatocytes, Biochem. Biophys. Res. Commun.90:587–595, 1979CrossRefPubMedGoogle Scholar
  17. 17.
    Keil, B.: Some newly characterized collagenases from procaryotes and lower eucaryotes, Mol. Cell. Biochem.23:87–108, 1979CrossRefPubMedGoogle Scholar
  18. 18.
    Lecroisey, A., Keil, B.: Differences in the degradation of native collagen by two microbial collagenases, Biochem. J.179:53–58, 1979PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1981

Authors and Affiliations

  • Jaro Sodek
    • 1
  • Johan N. M. Heersche
    • 1
  1. 1.Medical Research Council Group in Periodontal Physiology, Faculty of DentistryUniversity of TorontoTorontoCanada

Personalised recommendations