Search tips
Search criteria 


Logo of jcinvestThe Journal of Clinical Investigation
J Clin Invest. 1988 August; 82(2): 706–711.
PMCID: PMC303567

Cleavage and inactivation of alpha 1-antitrypsin by metalloproteinases released from neutrophils.


Human neutrophils, when stimulated with phorbol myristate acetate or fMet-Leu-Phe in the presence or absence of cytochalasin B, released metalloproteinases that catalytically inactivated the plasma serine proteinase inhibitor, alpha 1-antitrypsin. Inactivation, measured as loss of elastase inhibitory capacity, was accompanied by cleavage of a Mr 4,000 peptide from the COOH-terminus. Cleavage of alpha 1-antitrypsin by cell supernatants was inhibited by EDTA, o-phenanthroline, and DTT, but not by inhibitors of serine or thiol proteinases. Gelatinase and collagenase were separated from the medium of stimulated neutrophils. Both preparations cleaved and inactivated alpha 1-antitrypsin, with cleavage occurring close to the reactive center, at the Phe-Leu bond between positions P7 and P6. Cleavage by purified gelatinase was very slow and could account for only a minor fraction of the activity of neutrophil supernatants. The collagenase preparation was more active. However, the unusual cleavage site, and the ability of fMet-Leu-Phe-stimulated neutrophils to cleave alpha 1-antitrypsin without releasing collagenase, suggests that collagenase is not responsible for cleavage by the cells, which, by implication, is due to an as yet uncharacterized metalloenzyme. Our results demonstrate that by releasing metalloproteinases, neutrophils could proteolytically inactivate alpha 1-antitrypsin at sites of inflammation. This provides an alternative to the previously documented mechanism of inactivation by neutrophil-derived oxidants.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.3M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Dewald B, Rindler-Ludwig R, Bretz U, Baggiolini M. Subcellular localization and heterogeneity of neutral proteases in neutrophilic polymorphonuclear leukocytes. J Exp Med. 1975 Apr 1;141(4):709–723. [PMC free article] [PubMed]
  • Malemud CJ, Janoff A. Identification of neutral proteases in human neutrophil granules that degrade articular cartilage proteoglycan. Arthritis Rheum. 1975 Jul-Aug;18(4):361–368. [PubMed]
  • Baici A, Salgam P, Cohen G, Fehr K, Böni A. Action of collagenase and elastase from human polymorphonuclear leukocytes on human articular cartilage. Rheumatol Int. 1982;2(1):11–16. [PubMed]
  • Travis J, Salvesen GS. Human plasma proteinase inhibitors. Annu Rev Biochem. 1983;52:655–709. [PubMed]
  • Clark RA, Stone PJ, El Hag A, Calore JD, Franzblau C. Myeloperoxidase-catalyzed inactivation of alpha 1-protease inhibitor by human neutrophils. J Biol Chem. 1981 Apr 10;256(7):3348–3353. [PubMed]
  • Carp H, Miller F, Hoidal JR, Janoff A. Potential mechanism of emphysema: alpha 1-proteinase inhibitor recovered from lungs of cigarette smokers contains oxidized methionine and has decreased elastase inhibitory capacity. Proc Natl Acad Sci U S A. 1982 Mar;79(6):2041–2045. [PubMed]
  • Johnson D, Travis J. The oxidative inactivation of human alpha-1-proteinase inhibitor. Further evidence for methionine at the reactive center. J Biol Chem. 1979 May 25;254(10):4022–4026. [PubMed]
  • George PM, Vissers MC, Travis J, Winterbourn CC, Carrell RW. A genetically engineered mutant of alpha 1-antitrypsin protects connective tissue from neutrophil damage and may be useful in lung disease. Lancet. 1984 Dec 22;2(8417-8418):1426–1428. [PubMed]
  • Weiss SJ, Regiani S. Neutrophils degrade subendothelial matrices in the presence of alpha-1-proteinase inhibitor. Cooperative use of lysosomal proteinases and oxygen metabolites. J Clin Invest. 1984 May;73(5):1297–1303. [PMC free article] [PubMed]
  • Murphy G, Reynolds JJ, Bretz U, Baggiolini M. Partial purification of collagenase and gelatinase from human polymorphonuclear leucocytes. Analysis of their actions on soluble and insoluble collagens. Biochem J. 1982 Apr 1;203(1):209–221. [PubMed]
  • Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968;97:77–89. [PubMed]
  • Hibbs MS, Hasty KA, Seyer JM, Kang AH, Mainardi CL. Biochemical and immunological characterization of the secreted forms of human neutrophil gelatinase. J Biol Chem. 1985 Feb 25;260(4):2493–2500. [PubMed]
  • Murphy G, Bretz U, Baggiolini M, Reynolds JJ. The latent collagenase and gelatinase of human polymorphonuclear neutrophil leucocytes. Biochem J. 1980 Nov 15;192(2):517–525. [PubMed]
  • Hasty KA, Hibbs MS, Kang AH, Mainardi CL. Secreted forms of human neutrophil collagenase. J Biol Chem. 1986 Apr 25;261(12):5645–5650. [PubMed]
  • Sopata I. Further purification and some properties of a gelatin-specific proteinase of human leucocytes. Biochim Biophys Acta. 1982 Jul 16;717(1):26–31. [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • Laurell CB, Pierce J, Persson U, Thulin E. Purification of alpha1-antitrypsin from plasma through thiol-disulfide interchange. Eur J Biochem. 1975 Sep 1;57(1):107–113. [PubMed]
  • Bieth J, Spiess B, Wermuth CG. The synthesis and analytical use of a highly sensitive and convenient substrate of elastase. Biochem Med. 1974 Dec;11(4):350–357. [PubMed]
  • Carlson J, Stenflo J. The biosynthesis of rat alpha 1-antitrypsin. J Biol Chem. 1982 Nov 10;257(21):12987–12994. [PubMed]
  • Brennan SO, Carrell RW. alpha 1-Antitrypsin Christchurch, 363 Glu----Lys: mutation at the P'5 position does not affect inhibitory activity. Biochim Biophys Acta. 1986 Sep 5;873(1):13–19. [PubMed]
  • Tack BF, Dean J, Eilat D, Lorenz PE, Schechter AN. Tritium labeling of proteins to high specific radioactivity by reduction methylation. J Biol Chem. 1980 Sep 25;255(18):8842–8847. [PubMed]
  • Dewald B, Bretz U, Baggiolini M. Release of gelatinase from a novel secretory compartment of human neutrophils. J Clin Invest. 1982 Sep;70(3):518–525. [PMC free article] [PubMed]
  • Vissers MC, Winterbourn CC. Activation of human neutrophil gelatinase by endogenous serine proteinases. Biochem J. 1988 Jan 15;249(2):327–331. [PubMed]
  • Johnson D, Travis J. Inactivation of human alpha 1-proteinase inhibitor by thiol proteinases. Biochem J. 1977 Jun 1;163(3):639–641. [PubMed]
  • Virca GD, Lyerly D, Kreger A, Travis J. Inactivation of human plasma alpha 1-proteinase inhibitor by a metalloproteinase from Serratia marcescens. Biochim Biophys Acta. 1982 Jun 4;704(2):267–271. [PubMed]
  • Potempa J, Watorek W, Travis J. The inactivation of human plasma alpha 1-proteinase inhibitor by proteinases from Staphylococcus aureus. J Biol Chem. 1986 Oct 25;261(30):14330–14334. [PubMed]
  • Banda MJ, Clark EJ, Werb Z. Limited proteolysis by macrophage elastase inactivates human alpha 1-proteinase inhibitor. J Exp Med. 1980 Dec 1;152(6):1563–1570. [PMC free article] [PubMed]
  • Banda MJ, Clark EJ, Werb Z. Regulation of alpha 1 proteinase inhibitor function by rabbit alveolar macrophages. Evidence for proteolytic rather than oxidative inactivation. J Clin Invest. 1985 Jun;75(6):1758–1762. [PMC free article] [PubMed]
  • Boswell DR, Jeppsson JO, Brennan SO, Carrell RW. The reactive site of alpha 1-antitrypsin is C-terminal, not N-terminal. Biochim Biophys Acta. 1983 Apr 28;744(2):212–218. [PubMed]
  • Carp H, Janoff A. Potential mediator of inflammation. Phagocyte-derived oxidants suppress the elastase-inhibitory capacity of alpha 1-proteinase inhibitor in vitro. J Clin Invest. 1980 Nov;66(5):987–995. [PMC free article] [PubMed]
  • Winterbourn CC. Comparative reactivities of various biological compounds with myeloperoxidase-hydrogen peroxide-chloride, and similarity of the oxidant to hypochlorite. Biochim Biophys Acta. 1985 Jun 18;840(2):204–210. [PubMed]
  • Brot N, Fliss H, Coleman T, Weissbach H. Enzymatic reduction of methionine sulfoxide residues in proteins and peptides. Methods Enzymol. 1984;107:352–360. [PubMed]

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