[go: up one dir, main page]
More Web Proxy on the site http://driver.im/ Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1974 Apr;53(4):1044–1053. doi: 10.1172/JCI107641

Neutral Proteases and Cathepsin D in Human Articular Cartilage

Asher I Sapolsky 1,2,3, David S Howell 1,2,3, J Frederick Woessner Jr 1,2,3
PMCID: PMC333089  PMID: 4273625

Abstract

Proteolytic enzymes have been studied in extracts of human articular cartilage by the use of micromethods. The digestion of hemoglobin at pH 3.2 and of cartilage proteoglycan at pH 5 was shown to be due chiefly to cathepsin D. Cathepsin D was purified 900-fold from human patellar cartilage. Its identity was established by its specific cleavage of the B chain of insulin. At least six multiple forms of cathepsin D are present in cartilage; these corresponded to bovine forms 4-9. Cathepsin D had no action on proteins at pH 7.4. However, cartilage extracts digested proteoglycan, casein, and histone at this pH. The proteolytic activities against these three substrates were purified about 170-, 160-, and 70-fold, respectively. Each activity appeared in multiple forms on DEAE-Sephadex chromatography. The three activities appear to be different since cysteine inhibited casein digestion, aurothiomalate inhibited histone digestion, and neither inhibited proteoglycan digestion. Tests with a wide range of inhibitors and activators suggest that these three activities differ from other neutral proteases described in the literature.

Full text

PDF
1044

Images in this article

1048Image
on p.1048

Selected References

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

  1. Ali S. Y., Evans L. Enzymic degradation of cartilage in osteoarthritis. Fed Proc. 1973 Apr;32(4):1494–1498. [PubMed] [Google Scholar]
  2. Anderson A. J. Enzyme system in rat leucocyte granules which degrades insoluble collagen. Ann Rheum Dis. 1971 May;30(3):299–302. doi: 10.1136/ard.30.3.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BITTER T., MUIR H. M. A modified uronic acid carbazole reaction. Anal Biochem. 1962 Oct;4:330–334. doi: 10.1016/0003-2697(62)90095-7. [DOI] [PubMed] [Google Scholar]
  4. BOLLET A. J., HANDY J. R., STURGILL B. C. Chondroitin sulfate concentration and protein-polysaccharide composition of articular cartilage in osteoarthritis. J Clin Invest. 1963 Jun;42:853–859. doi: 10.1172/JCI104777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chrambach A., Reisfeld R. A., Wyckoff M., Zaccari J. A procedure for rapid and sensitive staining of protein fractionated by polyacrylamide gel electrophoresis. Anal Biochem. 1967 Jul;20(1):150–154. doi: 10.1016/0003-2697(67)90272-2. [DOI] [PubMed] [Google Scholar]
  6. Cowey F. K., Whitehouse M. W. Biochemical properties of anti-inflammatory drugs. VII. Inhibition of proteolytic enzymes in connective tissue by chloroquine (resochin) and related antimalarial antirheumatic drugs. Biochem Pharmacol. 1966 Aug;15(8):1071–1084. doi: 10.1016/0006-2952(66)90272-3. [DOI] [PubMed] [Google Scholar]
  7. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  8. Davies P., Krakauer K., Weissmann G. Calf thymus histone as a substrate for neutral and acid proteases of leucocyte lysosomes and other proteolytic enzymes. Anal Biochem. 1972 Feb;45(2):428–440. doi: 10.1016/0003-2697(72)90204-7. [DOI] [PubMed] [Google Scholar]
  9. Dingle J. T., Barrett A. J., Poole A. R., Stovin P. Inhibition by pepstatin of human cartilage degradation. Biochem J. 1972 Apr;127(2):443–444. doi: 10.1042/bj1270443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dingle J. T., Barrett A. J., Weston P. D. Cathepsin D. Characteristics of immunoinhibition and the confirmation of a role in cartilage breakdown. Biochem J. 1971 Jun;123(1):1–13. doi: 10.1042/bj1230001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Evanson J. M., Jeffrey J. J., Krane S. M. Studies on collagenase from rheumatoid synovium in tissue culture. J Clin Invest. 1968 Dec;47(12):2639–2651. doi: 10.1172/JCI105947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. FELL H. B., DINGLE J. T. Studies on the mode of action of excess of vitamin A. 6. Lysosomal protease and the degradation of cartilage matrix. Biochem J. 1963 May;87:403–408. doi: 10.1042/bj0870403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. FESSEL J. M., CHRISMAN O. D. ENZYMATIC DEGRADATION OF CHONDROMUCOPROTEIN BY CELL-FREE EXTRACTS OF HUMAN CARTILAGE. Arthritis Rheum. 1964 Aug;7:398–405. doi: 10.1002/art.1780070406. [DOI] [PubMed] [Google Scholar]
  14. Harris E. D., Jr, DiBona D. R., Krane S. M. Collagenases in human synovial fluid. J Clin Invest. 1969 Nov;48(11):2104–2113. doi: 10.1172/JCI106177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Hirschman A., Hirschman M. Aminopeptidase profile and protease activity in rat cartilage at physiological pH. Isr J Med Sci. 1971 Mar;7(3):403–405. [PubMed] [Google Scholar]
  17. Janoff A., Zeligs J. D. Vascular injury and lysis of basement membrane in vitro by neutral protease of human leukocytes. Science. 1968 Aug 16;161(3842):702–704. doi: 10.1126/science.161.3842.702. [DOI] [PubMed] [Google Scholar]
  18. Morishima H., Takita T., Aoyagi T., Takeuchi T., Umezawa H. The structure of pepstatin. J Antibiot (Tokyo) 1970 May;23(5):263–265. doi: 10.7164/antibiotics.23.263. [DOI] [PubMed] [Google Scholar]
  19. Morrison R. I., Barrett A. J., Dingle J. T., Prior D. Cathepsins BI and D. Action on human cartilage proteoglycans. Biochim Biophys Acta. 1973 Apr 12;302(2):411–419. doi: 10.1016/0005-2744(73)90170-8. [DOI] [PubMed] [Google Scholar]
  20. Moskowitz R. W., Klein L., Mast W. A. Current concepts of degenerative joint disease (osteoarthritis). Bull Rheum Dis. 1967 May;17(9):459–464. [PubMed] [Google Scholar]
  21. ORNSTEIN L. DISC ELECTROPHORESIS. I. BACKGROUND AND THEORY. Ann N Y Acad Sci. 1964 Dec 28;121:321–349. doi: 10.1111/j.1749-6632.1964.tb14207.x. [DOI] [PubMed] [Google Scholar]
  22. PRESS E. M., PORTER R. R., CEBRA J. The isolation and properties of a proteolytic enzyme, cathepsin D, from bovine spleen. Biochem J. 1960 Mar;74:501–514. doi: 10.1042/bj0740501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pita J. C., Müller F. J. Ultracentrifugal studies in capillary cells. I. Determination of sedimentation coefficients. Anal Biochem. 1972 Jun;47(2):395–407. doi: 10.1016/0003-2697(72)90133-9. [DOI] [PubMed] [Google Scholar]
  24. Rosenberg L., Pal S., Beale R., Schubert M. A comparison of proteinpolysaccharides of bovine nasal cartilage isolated and fractionated by different methods. J Biol Chem. 1970 Aug 25;245(16):4112–4122. [PubMed] [Google Scholar]
  25. Sapolsky A. I., Altman R. D., Howell D. S. Cathepsin D activity in normal and osteoarthritic human cartilage. Fed Proc. 1973 Apr;32(4):1489–1493. [PubMed] [Google Scholar]
  26. Sapolsky A. I., Altman R. D., Woessner J. F., Howell D. S. The action of cathepsin D in human articular cartilage on proteoglycans. J Clin Invest. 1973 Mar;52(3):624–633. doi: 10.1172/JCI107224. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sapolsky A. I., Woessner J. F., Jr Multiple forms of cathepsin D from bovine uterus. J Biol Chem. 1972 Apr 10;247(7):2069–2076. [PubMed] [Google Scholar]
  28. Wegelius O., Klockars M., Vainio K. Content of fibrocartilagenolytic enzymes and viscosity of homogenates of joint menisci in rheumatoid arthritis. Scand J Clin Lab Invest. 1970 Jan;25(1):41–45. doi: 10.3109/00365517009046188. [DOI] [PubMed] [Google Scholar]
  29. Weissmann G. Lysosomal mechanisms of tissue injury in arthritis. N Engl J Med. 1972 Jan 20;286(3):141–147. doi: 10.1056/NEJM197201202860307. [DOI] [PubMed] [Google Scholar]
  30. Weissmann G., Spilberg I., Krakauer K. Arthritis induced in rabbits by lysates of granulocyte lysosomes. Arthritis Rheum. 1969 Apr;12(2):103–116. doi: 10.1002/art.1780120207. [DOI] [PubMed] [Google Scholar]
  31. Woessner J. F., Jr Cartilage cathepsin D and its action on matrix components. Fed Proc. 1973 Apr;32(4):1485–1488. [PubMed] [Google Scholar]
  32. Woessner J. F., Jr Pepstatin inhibits the digestion of hemoglobin and protein-polysaccharide complex by cathepsin D. Biochem Biophys Res Commun. 1972 May 26;47(4):965–970. doi: 10.1016/0006-291x(72)90587-6. [DOI] [PubMed] [Google Scholar]
  33. Woessner J. F., Jr Purification of cathepsin D from cartilage and uterus and its action on the protein-polysaccharide complex of cartilage. J Biol Chem. 1973 Mar 10;248(5):1634–1642. [PubMed] [Google Scholar]
  34. Woessner J. F., Jr, Shamberger R. J., Jr Purification and properties of cathepsin D from bovine utrus. J Biol Chem. 1971 Apr 10;246(7):1951–1960. [PubMed] [Google Scholar]
  35. Wood G. C., Pryce-Jones R. H., White D. D., Nuki G. Chondromucoprotein-degrading neutral protease activity in rheumatoid synovial fluid. Ann Rheum Dis. 1971 Jan;30(1):73–77. doi: 10.1136/ard.30.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES