Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1980 Aug 1;189(2):349–357. doi: 10.1042/bj1890349

Properties of pig synovial collagenase.

J A Tyler, T E Cawston
PMCID: PMC1162004  PMID: 6257233

Abstract

1. Properties of a purified chemically activated form of pig synovial collagenase were examined and compared with a spontaneously active form of the enzyme. 2. The active enzyme has a specific activity of 53 000 units (microgram/min)/mg, a mol.wt. of 44 000 (by sodium dodecyl sulphate/polyarcylamide-gel electrophoresis in 2-mercaptoethanol) and pI 5.2 (by isoelectric focusing in polyacrylamide gels). 3. The activity has the characteristics of a metalloproteinase that degrades types I and III soluble or insoluble collagens in preference to type II, at an optimum pH of 6.5-8.5. 4. There is no detectable difference in these properties between the chemically activated and spontaneously active form of collagenase.

Full text

PDF
349

Images in this article

351Fig. 1.
on p.351
354Fig. 5.
on p.354
354Fig. 6.
on p.354

Selected References

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

  1. Banker G. A., Cotman C. W. Measurement of free electrophoretic mobility and retardation coefficient of protein-sodium dodecyl sulfate complexes by gel electrophoresis. A method to validate molecular weight estimates. J Biol Chem. 1972 Sep 25;247(18):5856–5861. [PubMed] [Google Scholar]
  2. Berman M. B., Manabe R. Corneal collagenases: evidence for zinc metalloenzymes. Ann Ophthalmol. 1973 Nov;5(11):1193–passim. [PubMed] [Google Scholar]
  3. Birkedal-Hansen H., Cobb C. M., Taylor R. E., Fullmer H. M. Activation of latent bovine gingival collagenase. Arch Oral Biol. 1975 Oct;20(10):681–685. doi: 10.1016/0003-9969(75)90137-5. [DOI] [PubMed] [Google Scholar]
  4. Burleigh M. C., Barrett A. J., Lazarus G. S. Cathepsin B1. A lysosomal enzyme that degrades native collagen. Biochem J. 1974 Feb;137(2):387–398. doi: 10.1042/bj1370387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cawston T. E., Barrett A. J. A rapid and reproducible assay for collagenase using [1-14C]acetylated collagen. Anal Biochem. 1979 Nov 1;99(2):340–345. doi: 10.1016/s0003-2697(79)80017-2. [DOI] [PubMed] [Google Scholar]
  6. Cawston T. E., Tyler J. A. Purification of pig synovial collagenase to high specific activity. Biochem J. 1979 Dec 1;183(3):647–656. doi: 10.1042/bj1830647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dingle J. T., Horsfield P., Fell H. B., Barratt M. E. Breakdown of proteoglycan and collagen induced in pig articular cartilage in organ culture. Ann Rheum Dis. 1975 Aug;34(4):303–311. doi: 10.1136/ard.34.4.303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dunker A. K., Kenyon A. J. Mobility of sodium dodecyl sulphate - protein complexes. Biochem J. 1976 Feb 1;153(2):191–197. doi: 10.1042/bj1530191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fiedler-Nagy C., Coffey J. W., Salvador R. A. Factors influencing the apparent molecular weight of collagenase produced by human-skin explants. Eur J Biochem. 1977 Jun 1;76(1):291–297. doi: 10.1111/j.1432-1033.1977.tb11595.x. [DOI] [PubMed] [Google Scholar]
  10. Gillet C., Eeckhout Y., Vaes G. Purification of procollagenase and collagenase by affinity chromatography on Sepharose-collagen. FEBS Lett. 1977 Feb 15;74(1):126–128. doi: 10.1016/0014-5793(77)80768-0. [DOI] [PubMed] [Google Scholar]
  11. Gross J. Collagen biology: structure, degradation, and disease. Harvey Lect. 1974;68:351–432. [PubMed] [Google Scholar]
  12. Harper E., Bloch K. J., Gross J. The zymogen of tadpole collagenase. Biochemistry. 1971 Aug 3;10(16):3035–3041. doi: 10.1021/bi00792a008. [DOI] [PubMed] [Google Scholar]
  13. Horwitz A. L., Hance A. J., Crystal R. G. Granulocyte collagenase: selective digestion of type I relative to type III collagen. Proc Natl Acad Sci U S A. 1977 Mar;74(3):897–901. doi: 10.1073/pnas.74.3.897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Liotta L. A., Abe S., Robey P. G., Martin G. R. Preferential digestion of basement membrane collagen by an enzyme derived from a metastatic murine tumor. Proc Natl Acad Sci U S A. 1979 May;76(5):2268–2272. doi: 10.1073/pnas.76.5.2268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McCroskery P. A., Richards J. F., Harris E. D., Jr Purification and characterization of a collagenase extracted from rabbit tumours. Biochem J. 1975 Oct;152(1):131–142. doi: 10.1042/bj1520131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Miller E. J., Harris E. D., Jr, Chung E., Finch J. E., Jr, McCroskery P. A., Butler W. T. Cleavage of Type II and III collagens with mammalian collagenase: site of cleavage and primary structure at the NH2-terminal portion of the smaller fragment released from both collagens. Biochemistry. 1976 Feb 24;15(4):787–792. doi: 10.1021/bi00649a009. [DOI] [PubMed] [Google Scholar]
  17. Nagai Y., Hori H. Entrapment of collagen in a polyacrylamide matrix and its application in the purification of animal collagenases. Biochim Biophys Acta. 1972 May 18;263(3):564–573. doi: 10.1016/0005-2795(72)90037-2. [DOI] [PubMed] [Google Scholar]
  18. Nagai Y., Lapiere C. M., Gross J. Tadpole collagenase. Preparation and purification. Biochemistry. 1966 Oct;5(10):3123–3130. doi: 10.1021/bi00874a007. [DOI] [PubMed] [Google Scholar]
  19. Ohyama H., Hashimoto K. Collagenase of human skin basal cell epithelioma. J Biochem. 1977 Jul;82(1):175–183. doi: 10.1093/oxfordjournals.jbchem.a131667. [DOI] [PubMed] [Google Scholar]
  20. Panyim S., Chalkley R. The molecular weights of vertebrate histones exploiting a modified sodium dodecyl sulfate electrophoretic method. J Biol Chem. 1971 Dec 25;246(24):7557–7560. [PubMed] [Google Scholar]
  21. Robertson P. B., Miller E. J. Cartilage collagen: inability to serve as a substrate for collagenases active against skin and bone collagen. Biochim Biophys Acta. 1972 Nov 10;289(1):247–250. doi: 10.1016/0005-2744(72)90129-5. [DOI] [PubMed] [Google Scholar]
  22. Sakamoto S., Sakamoto M., Matsumoto A., Goldhaber P., Glimcher M. J. Latent collagenase from the culture medium of embryonic chick bones. FEBS Lett. 1978 Apr 1;88(1):53–58. doi: 10.1016/0014-5793(78)80605-x. [DOI] [PubMed] [Google Scholar]
  23. Sellers A., Cartwright E., Murphy G., Reynolds J. J. Evidence that latent collagenases are enzyme-inhibitor complexes. Biochem J. 1977 May 1;163(2):303–307. doi: 10.1042/bj1630303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Seltzer J. L., Jeffrey J. J., Eisen A. Z. Evidence for mammalian collagenases as zinc ion metalloenzymes. Biochim Biophys Acta. 1977 Nov 23;485(1):179–187. doi: 10.1016/0005-2744(77)90205-4. [DOI] [PubMed] [Google Scholar]
  25. Seltzer J. L., Welgus H. G., Jeffrey J. J., Eisen A. Z. The function of Ca+ in the action of mammalian collagenases. Arch Biochem Biophys. 1976 Mar;173(1):355–361. doi: 10.1016/0003-9861(76)90270-8. [DOI] [PubMed] [Google Scholar]
  26. Shinkai H., Nagai Y. A latent collagenase from embryonic human skin explants. J Biochem. 1977 May;81(5):1261–1268. [PubMed] [Google Scholar]
  27. Shirahama K., Tsujii K., Takagi T. Free-boundary electrophoresis of sodium dodecyl sulfate-protein polypeptide complexes with special reference to SDS-polyacrylamide gel electrophoresis. J Biochem. 1974 Feb;75(2):309–319. doi: 10.1093/oxfordjournals.jbchem.a130398. [DOI] [PubMed] [Google Scholar]
  28. Stricklin G. P., Bauer E. A., Jeffrey J. J., Eisen A. Z. Human skin collagenase: isolation of precursor and active forms from both fibroblast and organ cultures. Biochemistry. 1977 Apr 19;16(8):1607–1615. doi: 10.1021/bi00627a013. [DOI] [PubMed] [Google Scholar]
  29. Stricklin G. P., Eisen A. Z., Bauer E. A., Jeffrey J. J. Human skin fibroblast collagenase: chemical properties of precursor and active forms. Biochemistry. 1978 Jun 13;17(12):2331–2337. doi: 10.1021/bi00605a012. [DOI] [PubMed] [Google Scholar]
  30. Vaes G. The release of collagenase as an inactive proenzyme by bone explants in culture. Biochem J. 1972 Jan;126(2):275–289. doi: 10.1042/bj1260275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Vater C. A., Mainardi C. L., Harris E. D., Jr Inhibitor of human collagenase from cultures of human tendon. J Biol Chem. 1979 Apr 25;254(8):3045–3053. [PubMed] [Google Scholar]
  32. Werb Z., Reynolds J. J. Purification and properties of a specific collagenase from rabbit synovial fibroblasts. Biochem J. 1975 Dec;151(3):645–653. doi: 10.1042/bj1510645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Woolley D. E., Glanville R. W., Crossley M. J., Evanson J. M. Purification of rheumatoid synovial collagenase and its action on soluble and insoluble collagen. Eur J Biochem. 1975 Jun;54(2):611–622. doi: 10.1111/j.1432-1033.1975.tb04173.x. [DOI] [PubMed] [Google Scholar]
  34. Woolley D. E., Glanville R. W., Evanson J. M. Differences in the physical properties of collagenases isolated from rheumatoid synovium and human skin. Biochem Biophys Res Commun. 1973 Apr 2;51(3):729–734. doi: 10.1016/0006-291x(73)91376-4. [DOI] [PubMed] [Google Scholar]
  35. Woolley D. E., Glanville R. W., Roberts D. R., Evanson J. M. Purification, characterization and inhibition of human skin collagenase. Biochem J. 1978 Feb 1;169(2):265–276. doi: 10.1042/bj1690265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Woolley D. E., Lindberg K. A., Glanville R. W., Evanson J. M. Action of rheumatoid synovial collagenase on cartilage collagen. Different susceptibilities of cartilage and tendon collagen to collagenase attack. Eur J Biochem. 1975 Jan 2;50(2):437–444. doi: 10.1111/j.1432-1033.1975.tb09821.x. [DOI] [PubMed] [Google Scholar]
  37. Woolley D. E., Roberts D. R., Evanson J. M. Small molecular weight beta 1 serum protein which specifically inhibits human collagenases. Nature. 1976 May 27;261(5558):325–327. doi: 10.1038/261325a0. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES