Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1985 Jul;49(1):52–60. doi: 10.1128/iai.49.1.52-60.1985

Subcellular localization and further characterization of a new elastase inhibitor from pneumococci.

M Vered, S R Simon, A Janoff
PMCID: PMC262057  PMID: 3847390

Abstract

Streptococcus pneumoniae contains an inhibitor of human neutrophil elastase. The agent does not inhibit other proteases, including neutrophil cathepsin G and pancreatic elastase. It is active in the presence of insoluble elastin as well as synthetic elastase substrates. The inhibitor is present in the pneumococcal cell membrane. [125I]elastase binding studies and inhibition experiments with intact bacterial autoplasts suggest that this agent has its elastase-binding site(s) exposed on the outside of the bacterial cell membrane. Native and randomized membrane vesicles also show equal inhibitory activity. Active inhibitor can be solubilized from pneumococcal membranes by treatment with a dipolar ionic detergent and can then be reconstituted, in active form, within artificial liposomes. Complex formation between the neutrophil elastase inhibitor and neutrophil elastase may involve noncovalent associations. Although elastase containing a covalently bound substrate analog no longer binds the pneumococcal inhibitor, the present study shows that complex formation is nevertheless independent of neutrophil elastase catalytic activity. Specific inhibitor activity and inhibitor release during bile salt-stimulated autolysis are greater in a nonnecrotizing pneumococcal strain (type I) than they are in a necrotizing strain (type III) or in Klebsiella pneumoniae. These results may help explain the frequent resolution of some pneumococcal pneumonias, despite the presence in the early pneumonic exudate of many neutrophils containing an elastolytic protease capable of injuring lung connective tissue.

Full text

PDF
52

Images in this article

58Image
on p.58

Selected References

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

  1. Bieth J., Spiess B., Wermuth C. G. The synthesis and analytical use of a highly sensitive and convenient substrate of elastase. Biochem Med. 1974 Dec;11(4):350–357. doi: 10.1016/0006-2944(74)90134-3. [DOI] [PubMed] [Google Scholar]
  2. Blondin J., Janoff A. The role of lysosomal elastase in the digestion of Escherichia coli proteins by human polymorphonuclear leukocytes: experiments with living leukocytes. J Clin Invest. 1976 Oct;58(4):971–979. doi: 10.1172/JCI108551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fritz H., Schiessler H., Gieger R., Ohlsson K., Hochstrasser K. Naturally occurring low molecular weight inhibitors of neutral proteinases from PMN-granulocytes and of kallikreins. Agents Actions. 1978 Jan;8(1-2):57–64. doi: 10.1007/BF01972403. [DOI] [PubMed] [Google Scholar]
  4. HUMMEL B. C. A modified spectrophotometric determination of chymotrypsin, trypsin, and thrombin. Can J Biochem Physiol. 1959 Dec;37:1393–1399. [PubMed] [Google Scholar]
  5. Hakenbeck R., Martin C., Morelli G. Streptococcus pneumoniae proteins released into medium upon inhibition of cell wall biosynthesis. J Bacteriol. 1983 Sep;155(3):1372–1381. doi: 10.1128/jb.155.3.1372-1381.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Henson P. M. Pathologic mechanisms in neutrophil-mediated injury. Am J Pathol. 1972 Sep;68(3):593–612. [PMC free article] [PubMed] [Google Scholar]
  7. Hjelmeland L. M., Chrambach A. Solubilization of functional membrane proteins. Methods Enzymol. 1984;104:305–318. doi: 10.1016/s0076-6879(84)04097-0. [DOI] [PubMed] [Google Scholar]
  8. Janoff A. Elastase in tissue injury. Annu Rev Med. 1985;36:207–216. doi: 10.1146/annurev.me.36.020185.001231. [DOI] [PubMed] [Google Scholar]
  9. Janoff A., Scherer J. Mediators of inflammation in leukocyte lysosomes. IX. Elastinolytic activity in granules of human polymorphonuclear leukocytes. J Exp Med. 1968 Nov 1;128(5):1137–1155. doi: 10.1084/jem.128.5.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Keiser H., Greenwald R. A., Feinstein G., Janoff A. Degradation of cartilage proteoglycan by human leukocyte granule neutral proteases--a model of joint injury. II. Degradation of isolated bovine nasal cartilage proteoglycan. J Clin Invest. 1976 Mar;57(3):625–632. doi: 10.1172/JCI108318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Lacks S., Neuberger M. Membrane location of a deoxyribonuclease implicated in the genetic transformation of Diplococcus pneumoniae. J Bacteriol. 1975 Dec;124(3):1321–1329. doi: 10.1128/jb.124.3.1321-1329.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Lonky S. A., Marsh J., Steele R., Jacobs K., Konopka R., Moser K. M. Protease and antiprotease responses in lung and peripheral blood in experimental canine pneumococcal pneumonia. Am Rev Respir Dis. 1980 Apr;121(4):685–693. doi: 10.1164/arrd.1980.121.4.685. [DOI] [PubMed] [Google Scholar]
  15. Mainardi C. L., Hasty D. L., Seyer J. M., Kang A. H. Specific cleavage of human type III collagen by human polymorphonuclear leukocyte elastase. J Biol Chem. 1980 Dec 25;255(24):12006–12010. [PubMed] [Google Scholar]
  16. McDonald J. A., Kelley D. G. Degradation of fibronectin by human leukocyte elastase. Release of biologically active fragments. J Biol Chem. 1980 Sep 25;255(18):8848–8858. [PubMed] [Google Scholar]
  17. McEnery M. W., Buhle E. L., Jr, Aebi U., Pedersen P. L. Proton ATPase of rat liver mitochondria. Preparation and visualization of a functional complex using the novel zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate. J Biol Chem. 1984 Apr 10;259(7):4642–4651. [PubMed] [Google Scholar]
  18. Odeberg H., Olsson I. Microbicidal mechanisms of human granulocytes: synergistic effects of granulocyte elastase and myeloperoxidase or chymotrypsin-like cationic protein. Infect Immun. 1976 Dec;14(6):1276–1283. doi: 10.1128/iai.14.6.1276-1283.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rabaud M., Dabadie P., Lefebvre F., Desgranges C., Bricaud H. Purification of human alpha 1 antiprotease-pancreatic elastase complex. Interaction with homologous elastin. Connect Tissue Res. 1984;12(2):165–174. doi: 10.3109/03008208408992781. [DOI] [PubMed] [Google Scholar]
  20. Suter S., Berger D., Despont J. P., Waldvogel F. Low levels of functional of alpha 1-proteinase inhibitor in the promotion of C3-cleavage by granulocyte-neutral proteases in pleural empyema. J Infect Dis. 1984 Jun;149(6):935–941. doi: 10.1093/infdis/149.6.935. [DOI] [PubMed] [Google Scholar]
  21. Tomasz A. Surface components of Streptococcus pneumoniae. Rev Infect Dis. 1981 Mar-Apr;3(2):190–211. doi: 10.1093/clinids/3.2.190. [DOI] [PubMed] [Google Scholar]
  22. Vered M., Dearing R., Janoff A. A new elastase inhibitor from Streptococcus pneumoniae protects against acute lung injury induced by neutrophil granules. Am Rev Respir Dis. 1985 Jan;131(1):131–133. doi: 10.1164/arrd.1985.131.1.131. [DOI] [PubMed] [Google Scholar]
  23. Vered M., Schutzbank T., Janoff A. Inhibitors of human neutrophil elastase in extracts of Streptococcus pneumoniae. Am Rev Respir Dis. 1984 Dec;130(6):1118–1124. doi: 10.1164/arrd.1984.130.6.1118. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES