Proteases II pp 115-122 | Cite as

Human Bronchial Proteinase Inhibitor: Rapid Purification Procedure and Inhibition of Luecocyte Elastase in Presence and in Absence of Human Lung Elastin

  • C. Boudier
  • D. Carvallo
  • M. Bruch
  • C. Roitsch
  • M. Courtney
  • J. G. Bieth
Conference paper
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 240)


Bronchial proteinase inhibitor (brl) is the most important inhibitor of human leucocyte elastase (HLE) present in upper airways secretions1. Its physiological function is probably to_ protect the bronchoalveolar tissues from HLE-induced proteolytic damage2,3 Published purification procedures of brl from bronchial sputum are based on Sepharose-bound chymotrypsin or trypsin, and yield an inhibitor preparation with a low specific activity and a multi-band pattern in SDS polyacrylamide gel electrophoresis4-6. In this paper we describe a purification procedure involving conventional chromatographic steps only and yielding a highly active inhibitor. The latter was used to study the kinetics of inhibition of HLE in a buffer system that mimics the physiological medium. Inhibition of HLE by brl was also investigated in presence of human lung elastin.


Secretory Leukocyte Protease Inhibitor Bronchial Secretion Inhibition Curve Leukocyte Elastase Human Seminal Plasma 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    H. Tegner, Quantitation of human granulocyte protease inhibitors in non-purulent bronchial lavage fluids. Acta Otolaryngol., 85: 282 (1978).PubMedGoogle Scholar
  2. 2.
    K. Ohlssori and H. Tegner, Inhibition of elastase from granulocytes by the low molecular weight bronchial protease inhibitor, Scand.J.Clin.Lab.Invest., 36: 437(1976).CrossRefGoogle Scholar
  3. 3.
    H. Schiessler, K. Hochstrasser and K. Ohlssori, Human mucous secretions biochemistry and possible biological Neutral Proteases of Human Poly-morphonuclear Leukocytes (Haveman, K. and Janoff, A., Eds), pp. 195 – 207, Urban & Schwarzenberg, Baltimore/Munich (1978).Google Scholar
  4. 4.
    K. Hochstrasser, K. Reichert, S. Schwarz, and E. Werle, Isolierung und Charakterisierung eines Proteaseninhibitors aus menschlichem Bronchialsekret, Hoppe Seyler’s Z.Physiol.Chem., 353: 221 (1972).PubMedCrossRefGoogle Scholar
  5. 5.
    K. Ohlsson, H. Tegner and U. Akesson, isolation and partial characterization of a low molecular weight acid stable protease inhibitor from human bronchial secretions. Hoppe-Seyler’s Z. Physiol. Chem., 358: 583(1977).PubMedCrossRefGoogle Scholar
  6. 6.
    C.E. Smith and D.A. Johnson, Human bronchial leucocyte proteinase inhibitor. Rapid isolation and kinetic analysis with human leucocyte proteinases, Biochem. J., 225: 463 (1985).PubMedGoogle Scholar
  7. 7.
    E.C. Klasen and J.A. Kramps, The N-terminal sequence of antileukoprotease isolated from bronchial secretion, Biochem. Biophys. Res. Commun., 128: 285(1985).PubMedCrossRefGoogle Scholar
  8. 8.
    U. Seemuler, M. Arnhold, H. Fritz, K. Wiedenmann, W. Machleidt, R. Heinzel, H. Appelhans, H-G. Gassen and F. Lottspeich, The acid-stable proteinase inhibitor of human mucous secretions (HUSH, anti-leukoprotease) Complete aminoacid sequence as revealed by protein and cDNA sequencing and structural homology to whey protein and red sea turtle proteinase inhibitor, FEBS Lett., 199: 43 (1986).CrossRefGoogle Scholar
  9. 9.
    R.C. Thompson and K. Ohlsson, Isolation, properties, and complete aminoacid sequence of human secretory leukocyte protease inhibitor, a potent inhibitor of leukocyte elastase, Proc. Natl. Acad. Sci. USA., 83: 6692(1986).PubMedCrossRefGoogle Scholar
  10. 10.
    C. Boudier, D. Carvallo, C. Roitsch, J.G. Bieth and M. Courtney, Purification and characterization of human bronchial proteinase inhibitor. Arch. Biochem. Biopys., 253: 439 (1987).CrossRefGoogle Scholar
  11. 11.
    C. Capellos and B. H. J. Bielski, Kinetic Systems. Mathematical description of chemical kinetics in solution, Krieger Pu. Co., Huntington, N.Y., USA (1980).Google Scholar
  12. 12.
    F. Gauthier, U. Fryksmark, K. Ohlsson, and J.G. Bieth, Kinetics of the inhibition of leucocyte elastase by the bronchial inhibitor, Biochim. Biophys. Acta,700: 178 (1982).PubMedCrossRefGoogle Scholar
  13. 13.
    J.G. Bieth, Pathophysiological interpretation of kinetic constants of protease inhibitors, Bull. Europ. Physiopath. resp.,16:(suppl.) 183 (1980).Google Scholar
  14. 14.
    J.G. Bieth, In vivo signifiance of kinetic constants of protein proteinase inhibitors. Biochem. Med., 32: 387 (1984).PubMedCrossRefGoogle Scholar
  15. 15.
    J.M. Tournier, J. Jacquot, P. Sadoul and J.G. Bieth, Non-competitive enzyme immunoassay for the measurement of bronchial inhibitor in biological fluids, Anal. Biochem., 131: 345 (1983).PubMedCrossRefGoogle Scholar
  16. 16.
    C. Boudier, A.Pelietier, A. Gast, J.M. Tournier, G. Pauli and J.G. Bieth, The elastase inhibitory capacity and the α1-proteinase inhibitor and bronchial inhibitor content of bronchoalveolar lavage fluids from healthy subjects, Biol. Chem. Hoppe-Seyler, in press (1987).Google Scholar
  17. 17.
    M.Bruch and J.G. Bieth, Influence of elastin on the inhibition of leucocyte elastase by α1-proteinase inhibitor and bronchial inhibitor, Biochem. J.,238: 269 (1986).PubMedGoogle Scholar
  18. 18.
    C.F. Reilly and J. Travis, The degradation of human lung elastin by neutrophil proteinases. Biochim. Biophys. Acta, 621:147 (1980).PubMedGoogle Scholar
  19. 19.
    W. Hornebeck and H. P. Schnebli, Effects of different elastase inhibitors on leukocyte elastase pre-adsorbed to elastin. Hoppe-Seyler’s Z. Physiol. Chem., 363: 455 (1982).PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • C. Boudier
    • 1
  • D. Carvallo
    • 2
  • M. Bruch
    • 1
  • C. Roitsch
    • 2
  • M. Courtney
    • 2
  • J. G. Bieth
    • 1
  1. 1.INSERM Unité 237Université Louis PasteurStrasbourgFrance
  2. 2.Transgène S.A. 11StrasbourgFrance

Personalised recommendations