Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1986 Jun;83(11):3756–3760. doi: 10.1073/pnas.83.11.3756

Human fibroblast collagenase: glycosylation and tissue-specific levels of enzyme synthesis.

S M Wilhelm, A Z Eisen, M Teter, S D Clark, A Kronberger, G Goldberg
PMCID: PMC323602  PMID: 3012533

Abstract

Human skin fibroblasts secrete collagenase as two proenzyme forms (57 and 52 kDa). The minor (57-kDa) proenzyme form is the result of a partial posttranslational modification of the major (52-kDa) proenzyme through the addition of N-linked complex oligosaccharides. Human endothelial cells as well as fibroblasts from human colon, cornea, gingiva, and lung also secrete collagenase in two forms indistinguishable from those of the skin fibroblast enzyme. In vitro tissue culture studies have shown that the level of constitutive synthesis of this fibroblast-type interstitial collagenase is tissue specific, varies widely, and correlates with the steady-state level of a single collagenase-specific mRNA of 2.5 kilobases. The tumor promoter, phorbol 12-myristate 13-acetate, apparently blocks the control of collagenase synthesis resulting in a similarly high level of collagenase expression (approximately equal to 3-7 micrograms of collagenase per 10(6) cells per 24 hr) in all examined cells. The constitutive level of synthesis of a 28-kDa collagenase inhibitor does not correlate with that of the enzyme. Phorbol 12-myristate 13-acetate stimulates the production of this inhibitor that in turn modulates the activity of collagenase in the conditioned media. As a result, the apparent activity of the enzyme present in the medium does not accurately reflect the rate of its synthesis and secretion.

Full text

PDF
3756

Images in this article

3757Image
on p.3757
3758Image
on p.3758
3758Image
on p.3758
3758Image
on p.3758
3759Image
on p.3759

Selected References

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

  1. Aggeler J., Frisch S. M., Werb Z. Collagenase is a major gene product of induced rabbit synovial fibroblasts. J Cell Biol. 1984 May;98(5):1656–1661. doi: 10.1083/jcb.98.5.1656. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bar-Shavit Z., Teitelbaum S. L., Stricklin G. P., Eisen A. Z., Kahn A. J., Welgus H. G. Differentiation of a human leukemia cell line and expression of collagenase inhibitor. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5380–5384. doi: 10.1073/pnas.82.16.5380. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bauer E. A., Cooper T. W., Huang J. S., Altman J., Deuel T. F. Stimulation of in vitro human skin collagenase expression by platelet-derived growth factor. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4132–4136. doi: 10.1073/pnas.82.12.4132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bauer E. A., Kronberger A., Stricklin G. P., Smith L. T., Holbrook K. A. Age-related changes in collagenase expression in cultured embryonic and fetal human skin fibroblasts. Exp Cell Res. 1985 Dec;161(2):484–494. doi: 10.1016/0014-4827(85)90103-x. [DOI] [PubMed] [Google Scholar]
  6. Bauer E. A., Stricklin G. P., Jeffrey J. J., Eisen A. Z. Collagenase production by human skin fibroblasts. Biochem Biophys Res Commun. 1975 May 5;64(1):232–240. doi: 10.1016/0006-291x(75)90243-0. [DOI] [PubMed] [Google Scholar]
  7. Brinckerhoff C. E., Benoit M. C., Culp W. J. Autoregulation of collagenase production by a protein synthesized and secreted by synovial fibroblasts: cellular mechanism for control of collagen degradation. Proc Natl Acad Sci U S A. 1985 Apr;82(7):1916–1920. doi: 10.1073/pnas.82.7.1916. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brinckerhoff C. E., Gross R. H., Nagase H., Sheldon L., Jackson R. C., Harris E. D., Jr Increased level of translatable collagenase messenger ribonucleic acid in rabbit synovial fibroblasts treated with phorbol myristate acetate or crystals of monosodium urate monohydrate. Biochemistry. 1982 May 25;21(11):2674–2679. doi: 10.1021/bi00540a015. [DOI] [PubMed] [Google Scholar]
  9. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  10. Cawston T. E., Murphy G., Mercer E., Galloway W. A., Hazleman B. L., Reynolds J. J. The interaction of purified rabbit bone collagenase with purified rabbit bone metalloproteinase inhibitor. Biochem J. 1983 May 1;211(2):313–318. doi: 10.1042/bj2110313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Christner P., Damato D., Reinhart M., Abrams W. Purification of human neutrophil collagenase and production of a monospecific antiserum. Biochemistry. 1982 Nov 9;21(23):6005–6011. doi: 10.1021/bi00266a043. [DOI] [PubMed] [Google Scholar]
  12. Chua C. C., Geiman D. E., Keller G. H., Ladda R. L. Induction of collagenase secretion in human fibroblast cultures by growth promoting factors. J Biol Chem. 1985 May 10;260(9):5213–5216. [PubMed] [Google Scholar]
  13. Clark S. D., Wilhelm S. M., Stricklin G. P., Welgus H. G. Coregulation of collagenase and collagenase inhibitor production by phorbol myristate acetate in human skin fibroblasts. Arch Biochem Biophys. 1985 Aug 15;241(1):36–44. doi: 10.1016/0003-9861(85)90358-3. [DOI] [PubMed] [Google Scholar]
  14. Cooper T. W., Bauer E. A., Eisen A. Z. Enzyme-linked immunosorbent assay for human skin collagenase. Coll Relat Res. 1983 May;3(3):205–215. doi: 10.1016/s0174-173x(83)80004-1. [DOI] [PubMed] [Google Scholar]
  15. Eaton D. L., Scott R. W., Baker J. B. Purification of human fibroblast urokinase proenzyme and analysis of its regulation by proteases and protease nexin. J Biol Chem. 1984 May 25;259(10):6241–6247. [PubMed] [Google Scholar]
  16. Elder J. H., Alexander S. endo-beta-N-acetylglucosaminidase F: endoglycosidase from Flavobacterium meningosepticum that cleaves both high-mannose and complex glycoproteins. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4540–4544. doi: 10.1073/pnas.79.15.4540. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gordon J. M., Bauer E. A., Eisen A. Z. Collagenase in human cornea: immunologic localization. Arch Ophthalmol. 1980 Feb;98(2):341–345. doi: 10.1001/archopht.1980.01020030337022. [DOI] [PubMed] [Google Scholar]
  18. Grillo H. C., Gross J. Collagenolytic activity during mammalian wound repair. Dev Biol. 1967 Apr;15(4):300–317. doi: 10.1016/0012-1606(67)90029-2. [DOI] [PubMed] [Google Scholar]
  19. Guan J. L., Machamer C. E., Rose J. K. Glycosylation allows cell-surface transport of an anchored secretory protein. Cell. 1985 Sep;42(2):489–496. doi: 10.1016/0092-8674(85)90106-0. [DOI] [PubMed] [Google Scholar]
  20. Harris E. D., Jr, Welgus H. G., Krane S. M. Regulation of the mammalian collagenases. Coll Relat Res. 1984 Dec;4(6):493–512. doi: 10.1016/s0174-173x(84)80015-1. [DOI] [PubMed] [Google Scholar]
  21. Hasty K. A., Hibbs M. S., Kang A. H., Mainardi C. L. Heterogeneity among human collagenases demonstrated by monoclonal antibody that selectively recognizes and inhibits human neutrophil collagenase. J Exp Med. 1984 May 1;159(5):1455–1463. doi: 10.1084/jem.159.5.1455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  24. Macartney H. W., Tschesche H. Latent and active human polymorphonuclear leukocyte collagenases. Isolation, purification and characterisation. Eur J Biochem. 1983 Jan 17;130(1):71–78. doi: 10.1111/j.1432-1033.1983.tb07118.x. [DOI] [PubMed] [Google Scholar]
  25. Matrisian L. M., Glaichenhaus N., Gesnel M. C., Breathnach R. Epidermal growth factor and oncogenes induce transcription of the same cellular mRNA in rat fibroblasts. EMBO J. 1985 Jun;4(6):1435–1440. doi: 10.1002/j.1460-2075.1985.tb03799.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Moscatelli D., Jaffe E., Rifkin D. B. Tetradecanoyl phorbol acetate stimulates latent collagenase production by cultured human endothelial cells. Cell. 1980 Jun;20(2):343–351. doi: 10.1016/0092-8674(80)90620-0. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Nagase H., Brinckerhoff C. E., Vater C. A., Harris E. D., Jr Biosynthesis and secretion of procollagenase by rabbit synovial fibroblasts. Inhibition of procollagenase secretion by monensin and evidence for glycosylation of procollagenase. Biochem J. 1983 Aug 15;214(2):281–288. doi: 10.1042/bj2140281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schliwa M., Nakamura T., Porter K. R., Euteneuer U. A tumor promoter induces rapid and coordinated reorganization of actin and vinculin in cultured cells. J Cell Biol. 1984 Sep;99(3):1045–1059. doi: 10.1083/jcb.99.3.1045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sperling R., Sperling J., Levine A. D., Spann P., Stark G. R., Kornberg R. D. Abundant nuclear ribonucleoprotein form of CAD RNA. Mol Cell Biol. 1985 Mar;5(3):569–575. doi: 10.1128/mcb.5.3.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. 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]
  33. Stricklin G. P., Jeffrey J. J., Roswit W. T., Eisen A. Z. Human skin fibroblast procollagenase: mechanisms of activation by organomercurials and trypsin. Biochemistry. 1983 Jan 4;22(1):61–68. doi: 10.1021/bi00270a009. [DOI] [PubMed] [Google Scholar]
  34. Struck D. K., Lennarz W. J. Evidence for the participation of saccharide-lipids in the synthesis of the oligosaccharide chain of ovalbumin. J Biol Chem. 1977 Feb 10;252(3):1007–1013. [PubMed] [Google Scholar]
  35. Tarentino A. L., Maley F. Purification and properties of an endo-beta-N-acetylglucosaminidase from Streptomyces griseus. J Biol Chem. 1974 Feb 10;249(3):811–817. [PubMed] [Google Scholar]
  36. Valle K. J., Bauer E. A. Biosynthesis of collagenase by human skin fibroblasts in monolayer culture. J Biol Chem. 1979 Oct 25;254(20):10115–10122. [PubMed] [Google Scholar]
  37. Vater C. A., Nagase H., Harris E. D., Jr Purification of an endogenous activator of procollagenase from rabbit synovial fibroblast culture medium. J Biol Chem. 1983 Aug 10;258(15):9374–9382. [PubMed] [Google Scholar]
  38. Welgus H. G., Campbell E. J., Bar-Shavit Z., Senior R. M., Teitelbaum S. L. Human alveolar macrophages produce a fibroblast-like collagenase and collagenase inhibitor. J Clin Invest. 1985 Jul;76(1):219–224. doi: 10.1172/JCI111949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Welgus H. G., Jeffrey J. J., Eisen A. Z., Roswit W. T., Stricklin G. P. Human skin fibroblast collagenase: interaction with substrate and inhibitor. Coll Relat Res. 1985 Mar;5(2):167–179. doi: 10.1016/s0174-173x(85)80038-8. [DOI] [PubMed] [Google Scholar]
  40. Welgus H. G., Jeffrey J. J., Eisen A. Z. The collagen substrate specificity of human skin fibroblast collagenase. J Biol Chem. 1981 Sep 25;256(18):9511–9515. [PubMed] [Google Scholar]
  41. Welgus H. G., Stricklin G. P., Eisen A. Z., Bauer E. A., Cooney R. V., Jeffrey J. J. A specific inhibitor of vertebrate collagenase produced by human skin fibroblasts. J Biol Chem. 1979 Mar 25;254(6):1938–1943. [PubMed] [Google Scholar]
  42. Welgus H. G., Stricklin G. P. Human skin fibroblast collagenase inhibitor. Comparative studies in human connective tissues, serum, and amniotic fluid. J Biol Chem. 1983 Oct 25;258(20):12259–12264. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES