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. 1989 Feb 15;258(1):115–119. doi: 10.1042/bj2580115

Purification of the neutral proteoglycan-degrading metalloproteinase from human articular cartilage tissue and its identification as stromelysin matrix metalloproteinase-3.

Z Gunja-Smith 1, H Nagase 1, J F Woessner Jr 1
PMCID: PMC1138331  PMID: 2930500

Abstract

The 'neutral' proteoglycan-degrading metalloproteinase of human articular cartilage was purified 3,500-fold by use of an anti-(matrix metalloproteinase-3) immunoglobulin G affinity column. Molecular masses of the latent and multiple active forms and specificity of action on casein, transferrin, gelatin and fibronectin were identical with those of authentic stromelysin (matrix metalloproteinase-3) from cultured human rheumatoid synovial fibroblasts. The optimum pH of this proteinase on proteoglycan monomer was pH 5.5, and on Azocoll, 6.2; digestion of fibronectin and gelatin was more extensive at pH 5.5 than at 7.5.

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Selected References

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

  1. Azzo W., Woessner J. F., Jr Purification and characterization of an acid metalloproteinase from human articular cartilage. J Biol Chem. 1986 Apr 25;261(12):5434–5441. [PubMed] [Google Scholar]
  2. Boucek R. J., Noble N. L., Gunja-Smith Z., Simpson C. F. Considerations of aortic elastin chemistry in the genesis of the intimal plaque (broad-breasted white turkey). Exp Mol Pathol. 1979 Dec;31(3):400–412. doi: 10.1016/0014-4800(79)90040-6. [DOI] [PubMed] [Google Scholar]
  3. Campbell I. K., Golds E. E., Mort J. S., Roughley P. J. Human articular cartilage secretes characteristic metal dependent proteinases upon stimulation by mononuclear cell factor. J Rheumatol. 1986 Feb;13(1):20–27. [PubMed] [Google Scholar]
  4. Chin J. R., Murphy G., Werb Z. Stromelysin, a connective tissue-degrading metalloendopeptidase secreted by stimulated rabbit synovial fibroblasts in parallel with collagenase. Biosynthesis, isolation, characterization, and substrates. J Biol Chem. 1985 Oct 5;260(22):12367–12376. [PubMed] [Google Scholar]
  5. Dean D. D., Woessner J. F., Jr A sensitive, specific assay for tissue collagenase using telopeptide-free [3H]acetylated collagen. Anal Biochem. 1985 Jul;148(1):174–181. doi: 10.1016/0003-2697(85)90642-6. [DOI] [PubMed] [Google Scholar]
  6. Galloway W. A., Murphy G., Sandy J. D., Gavrilovic J., Cawston T. E., Reynolds J. J. Purification and characterization of a rabbit bone metalloproteinase that degrades proteoglycan and other connective-tissue components. Biochem J. 1983 Mar 1;209(3):741–752. doi: 10.1042/bj2090741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ito A., Nagase H. Evidence that human rheumatoid synovial matrix metalloproteinase 3 is an endogenous activator of procollagenase. Arch Biochem Biophys. 1988 Nov 15;267(1):211–216. doi: 10.1016/0003-9861(88)90025-2. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Martel-Pelletier J., Pelletier J. P., Cloutier J. M., Howell D. S., Ghandur-Mnaymneh L., Woessner J. F., Jr Neutral proteases capable of proteoglycan digesting activity in osteoarthritic and normal human articular cartilage. Arthritis Rheum. 1984 Mar;27(3):305–312. doi: 10.1002/art.1780270310. [DOI] [PubMed] [Google Scholar]
  10. Murphy G., Cockett M. I., Stephens P. E., Smith B. J., Docherty A. J. Stromelysin is an activator of procollagenase. A study with natural and recombinant enzymes. Biochem J. 1987 Nov 15;248(1):265–268. doi: 10.1042/bj2480265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Murphy G., Nagase H., Brinckerhoff C. E. Relationship of procollagenase activator, stromelysin and matrix metalloproteinase 3. Coll Relat Res. 1988 Jul;8(4):389–391. doi: 10.1016/s0174-173x(88)80009-8. [DOI] [PubMed] [Google Scholar]
  12. Nagase H., Woessner J. F., Jr An improved assay for proteases and polysaccharidases employing a cartilage proteoglycan substrate entrapped in polyacrylamide particles. Anal Biochem. 1980 Sep 15;107(2):385–392. doi: 10.1016/0003-2697(80)90400-5. [DOI] [PubMed] [Google Scholar]
  13. Okada Y., Harris E. D., Jr, Nagase H. The precursor of a metalloendopeptidase from human rheumatoid synovial fibroblasts. Purification and mechanisms of activation by endopeptidases and 4-aminophenylmercuric acetate. Biochem J. 1988 Sep 15;254(3):731–741. doi: 10.1042/bj2540731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Okada Y., Nagase H., Harris E. D., Jr A metalloproteinase from human rheumatoid synovial fibroblasts that digests connective tissue matrix components. Purification and characterization. J Biol Chem. 1986 Oct 25;261(30):14245–14255. [PubMed] [Google Scholar]
  15. Sapolsky A. I., Howell D. S. Further characterization of a neutral metalloprotease isolated from human articular cartilage. Arthritis Rheum. 1982 Aug;25(8):981–988. doi: 10.1002/art.1780250811. [DOI] [PubMed] [Google Scholar]
  16. Sapolsky A. I., Keiser H., Howell D. S., Woessner J. F., Jr Metalloproteases of human articular cartilage that digest cartilage proteoglycan at neutral and acid pH. J Clin Invest. 1976 Oct;58(4):1030–1041. doi: 10.1172/JCI108526. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schwartz R. C., Greer C. L., Gegenheimer P., Abelson J. Enzymatic mechanism of an RNA ligase from wheat germ. J Biol Chem. 1983 Jul 10;258(13):8374–8383. [PubMed] [Google Scholar]
  18. Treadwell B. V., Neidel J., Pavia M., Towle C. A., Trice M. E., Mankin H. J. Purification and characterization of collagenase activator protein synthesized by articular cartilage. Arch Biochem Biophys. 1986 Dec;251(2):715–723. doi: 10.1016/0003-9861(86)90381-4. [DOI] [PubMed] [Google Scholar]
  19. Vater C. A., Hahn J. L., Harris E. D., Jr Preparation of a monospecific antibody to purified rabbit synovial fibroblast collagenase. Coll Relat Res. 1981 Nov;1(6):527–542. doi: 10.1016/s0174-173x(81)80034-9. [DOI] [PubMed] [Google Scholar]
  20. Wilhelm S. M., Collier I. E., Kronberger A., Eisen A. Z., Marmer B. L., Grant G. A., Bauer E. A., Goldberg G. I. Human skin fibroblast stromelysin: structure, glycosylation, substrate specificity, and differential expression in normal and tumorigenic cells. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6725–6729. doi: 10.1073/pnas.84.19.6725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Woessner J. F., Jr, Selzer M. G. Two latent metalloproteases of human articular cartilage that digest proteoglycan. J Biol Chem. 1984 Mar 25;259(6):3633–3638. [PubMed] [Google Scholar]
  22. Wray W., Boulikas T., Wray V. P., Hancock R. Silver staining of proteins in polyacrylamide gels. Anal Biochem. 1981 Nov 15;118(1):197–203. doi: 10.1016/0003-2697(81)90179-2. [DOI] [PubMed] [Google Scholar]

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