Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1997 May 1;323(Pt 3):719–725. doi: 10.1042/bj3230719

Sheep mast cell proteinase-1, a serine proteinase with both tryptase- and chymase-like properties, is inhibited by plasma proteinase inhibitors and is mitogenic for bovine pulmonary artery fibroblasts.

A D Pemberton 1, C M Belham 1, J F Huntley 1, R Plevin 1, H R Miller 1
PMCID: PMC1218375  PMID: 9169605

Abstract

Sheep mast cell proteinase-1 (sMCP-1), a serine proteinase with dual chymase/tryptase activity, is expressed in gastrointestinal mast cells, and released systemically and on to the mucosal surface during gastrointestinal nematode infection. The potential for native plasma proteinase inhibitors to control sMCP-1 activity was investigated. Sheep alpha1-proteinase inhibitor (alpha1PI) inhibited sMCP-1 slowly, with second-order association rate constant (kass) 1. 1x10(3) M-1.s-1, whereas sheep contrapsin inhibited trypsin (kass 2.2x10(6) M-1.s-1) but not sMCP-1. Western-blot analysis and gel filtration showed that when added to serum or plasma, sMCP-1 was partitioned between alpha1PI and alpha2-macroglobulin. The possibility that significant cleavage of plasma proteins could occur before sMCP-1 was inhibited was investigated using gel filtration and SDS/PAGE after adding sMCP-1 to plasma. Cleavage of ovine fibrinogen occurred in the presence of excess alpha1PI and alpha2-macroglobulin, the alpha-chain being cleaved C-terminally and the beta-chain at the putative Lys-27. In addition, sMCP-1 was found to be mitogenic for bovine pulmonary artery fibroblasts, but was not mitogenic in the presence of soya-bean trypsin inhibitor. In terms of fibrinogen cleavage and fibroblast stimulation, sMCP-1 shows functional similarities to mast cell tryptase.

Full Text

The Full Text of this article is available as a PDF (393.1 KB).

Selected References

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

  1. Boudier C., Bieth J. G. Mucus proteinase inhibitor: a fast-acting inhibitor of leucocyte elastase. Biochim Biophys Acta. 1989 Mar 16;995(1):36–41. doi: 10.1016/0167-4838(89)90230-6. [DOI] [PubMed] [Google Scholar]
  2. Caughey G. H., Viro N. F., Lazarus S. C., Nadel J. A. Purification and characterization of dog mastocytoma chymase: identification of an octapeptide conserved in chymotryptic leukocyte proteinases. Biochim Biophys Acta. 1988 Jan 29;952(2):142–149. doi: 10.1016/0167-4838(88)90109-4. [DOI] [PubMed] [Google Scholar]
  3. Christensen S., Sottrup-Jensen L. Characterization of two serpins from bovine plasma and milk. Biochem J. 1994 Oct 15;303(Pt 2):383–390. doi: 10.1042/bj3030383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chung D. W., Que B. G., Rixon M. W., Mace M., Jr, Davie E. W. Characterization of complementary deoxyribonucleic acid and genomic deoxyribonucleic acid for the beta chain of human fibrinogen. Biochemistry. 1983 Jun 21;22(13):3244–3250. doi: 10.1021/bi00282a032. [DOI] [PubMed] [Google Scholar]
  5. Chung D. W., Rixon M. W., MacGillivray R. T., Davie E. W. Characterization of a cDNA clone coding for the beta chain of bovine fibrinogen. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1466–1470. doi: 10.1073/pnas.78.3.1466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dubin A., Koj A., Chudzik J. Isolation and some molecular parameters of elastase-like normal proteinases from horse blood leucocytes. Biochem J. 1976 Feb 1;153(2):389–396. doi: 10.1042/bj1530389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Feinman R. D. The proteinase-binding reaction of alpha 2M. Ann N Y Acad Sci. 1994 Sep 10;737:245–266. doi: 10.1111/j.1749-6632.1994.tb44316.x. [DOI] [PubMed] [Google Scholar]
  8. Gibson S., Miller H. R. Mast cell subsets in the rat distinguished immunohistochemically by their content of serine proteinases. Immunology. 1986 May;58(1):101–104. [PMC free article] [PubMed] [Google Scholar]
  9. Gruber B. L., Marchese M. J., Suzuki K., Schwartz L. B., Okada Y., Nagase H., Ramamurthy N. S. Synovial procollagenase activation by human mast cell tryptase dependence upon matrix metalloproteinase 3 activation. J Clin Invest. 1989 Nov;84(5):1657–1662. doi: 10.1172/JCI114344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hill R. E., Hastie N. D. Accelerated evolution in the reactive centre regions of serine protease inhibitors. Nature. 1987 Mar 5;326(6108):96–99. doi: 10.1038/326096a0. [DOI] [PubMed] [Google Scholar]
  11. Huntley J. F., Gibson S., Knox D., Miller H. R. The isolation and purification of a proteinase with chymotrypsin-like properties from ovine mucosal mast cells. Int J Biochem. 1986;18(8):673–682. doi: 10.1016/0020-711x(86)90389-7. [DOI] [PubMed] [Google Scholar]
  12. Huntley J. F., Gooden C., Newlands G. F., Mackellar A., Lammas D. A., Wakelin D., Tuohy M., Woodbury R. G., Miller H. R. Distribution of intestinal mast cell proteinase in blood and tissues of normal and Trichinella-infected mice. Parasite Immunol. 1990 Jan;12(1):85–95. doi: 10.1111/j.1365-3024.1990.tb00938.x. [DOI] [PubMed] [Google Scholar]
  13. Huntley J. F., Newlands G. F., Jackson F., Miller H. R. The influence of challenge dose, duration of immunity, or steroid treatment on mucosal mast cells and on the distribution of sheep mast cell proteinase in Haemonchus-infected sheep. Parasite Immunol. 1992 Jul;14(4):429–440. doi: 10.1111/j.1365-3024.1992.tb00017.x. [DOI] [PubMed] [Google Scholar]
  14. Irvine J., Newlands G. F., Huntley J. F., Miller H. R. Interaction of murine intestinal mast cell proteinase with inhibitors (serpins) in blood; analysis by SDS-PAGE and western blotting. Immunology. 1990 Jan;69(1):139–144. [PMC free article] [PubMed] [Google Scholar]
  15. Jameson G. W., Roberts D. V., Adams R. W., Kyle W. S., Elmore D. T. Determination of the operational molarity of solutions of bovine alpha-chymotrypsin, trypsin, thrombin and factor Xa by spectrofluorimetric titration. Biochem J. 1973 Jan;131(1):107–117. doi: 10.1042/bj1310107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kan C. C., Solomon E., Belt K. T., Chain A. C., Hiorns L. R., Fey G. Nucleotide sequence of cDNA encoding human alpha 2-macroglobulin and assignment of the chromosomal locus. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2282–2286. doi: 10.1073/pnas.82.8.2282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mistry R., Snashall P. D., Totty N., Guz A., Tetley T. D. Isolation and characterization of sheep alpha 1-proteinase inhibitor. Biochem J. 1991 Feb 1;273(Pt 3):685–690. doi: 10.1042/bj2730685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pemberton A. D., Huntley J. F., Miller H. R. Sheep mast cell proteinase-1: characterization as a member of a new class of dual-specific ruminant chymases. Biochem J. 1997 Feb 1;321(Pt 3):665–670. doi: 10.1042/bj3210665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pemberton A. D., Miller H. R., John H. A., Scudamore C. L. Comparative studies of the Spi1 proteins of three equine alpha-1-proteinase inhibitor haplotypes following isolation by affinity chromatography. Int J Biochem. 1993 Sep;25(9):1263–1268. doi: 10.1016/0020-711x(93)90077-r. [DOI] [PubMed] [Google Scholar]
  20. Pirie-Shepherd S. R., Miller H. R., Ryle A. Differential inhibition of rat mast cell proteinase I and II by members of the alpha-1-proteinase inhibitor family of serine proteinase inhibitors. J Biol Chem. 1991 Sep 15;266(26):17314–17319. [PubMed] [Google Scholar]
  21. Potempa J., Enghild J. J., Travis J. The primary elastase inhibitor (elastasin) and trypsin inhibitor (contrapsin) in the goat are serpins related to human alpha 1-anti-chymotrypsin. Biochem J. 1995 Feb 15;306(Pt 1):191–197. doi: 10.1042/bj3060191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ruoss S. J., Hartmann T., Caughey G. H. Mast cell tryptase is a mitogen for cultured fibroblasts. J Clin Invest. 1991 Aug;88(2):493–499. doi: 10.1172/JCI115330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schechter N. M., Fräki J. E., Geesin J. C., Lazarus G. S. Human skin chymotryptic proteinase. Isolation and relation to cathepsin g and rat mast cell proteinase I. J Biol Chem. 1983 Mar 10;258(5):2973–2978. [PubMed] [Google Scholar]
  24. Schechter N. M., Jordan L. M., James A. M., Cooperman B. S., Wang Z. M., Rubin H. Reaction of human chymase with reactive site variants of alpha 1-antichymotrypsin. Modulation of inhibitor versus substrate properties. J Biol Chem. 1993 Nov 5;268(31):23626–23633. [PubMed] [Google Scholar]
  25. Schechter N. M., Sprows J. L., Schoenberger O. L., Lazarus G. S., Cooperman B. S., Rubin H. Reaction of human skin chymotrypsin-like proteinase chymase with plasma proteinase inhibitors. J Biol Chem. 1989 Dec 15;264(35):21308–21315. [PubMed] [Google Scholar]
  26. Schwartz L. B., Bradford T. R., Littman B. H., Wintroub B. U. The fibrinogenolytic activity of purified tryptase from human lung mast cells. J Immunol. 1985 Oct;135(4):2762–2767. [PubMed] [Google Scholar]
  27. Schwartz L. B. Tryptase: a mast cell serine protease. Methods Enzymol. 1994;244:88–100. doi: 10.1016/0076-6879(94)44008-5. [DOI] [PubMed] [Google Scholar]
  28. Schägger H., von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. doi: 10.1016/0003-2697(87)90587-2. [DOI] [PubMed] [Google Scholar]
  29. Scudamore C. L., Thornton E. M., McMillan L., Newlands G. F., Miller H. R. Release of the mucosal mast cell granule chymase, rat mast cell protease-II, during anaphylaxis is associated with the rapid development of paracellular permeability to macromolecules in rat jejunum. J Exp Med. 1995 Dec 1;182(6):1871–1881. doi: 10.1084/jem.182.6.1871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sinha U., Sinha S., Janoff A. Characterization of sheep alpha-1-proteinase inhibitor. Important differences from the human protein. Am Rev Respir Dis. 1988 Mar;137(3):558–563. doi: 10.1164/ajrccm/137.3.558. [DOI] [PubMed] [Google Scholar]
  31. Smith T. J., Hougland M. W., Johnson D. A. Human lung tryptase. Purification and characterization. J Biol Chem. 1984 Sep 10;259(17):11046–11051. [PubMed] [Google Scholar]
  32. Trong H. L., Newlands G. F., Miller H. R., Charbonneau H., Neurath H., Woodbury R. G. Amino acid sequence of a mouse mucosal mast cell protease. Biochemistry. 1989 Jan 10;28(1):391–395. doi: 10.1021/bi00427a054. [DOI] [PubMed] [Google Scholar]
  33. Woodbury R. G., Everitt M. T., Neurath H. Mast cell proteases. Methods Enzymol. 1981;80(Pt 100):588–609. doi: 10.1016/s0076-6879(81)80047-x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES