Human Leukocyte Elastase and Cathepsin G Inactivate Factor VII by Limited Proteolysis

 

PDF Download Buy Article Permissions and Reprints

Summary

The effect of supernatant from phorbol myristate acetate (PMA) stimulated human polymorphonuclear granulocytes (PMN) on human factor VII was studied in vitro. The supernatant caused a rapid loss in factor VII coagulant activity by the action of human leukocyte elastase (HLE) and cathepsin G in the supernatant, as demonstrated by the use of specific inhibitors of the two serine proteases, respectively. Preincubation of the supernatant with the elastase inhibitor and the cathepsin G inhibitor preserved 80% and 25% of the clotting activity, respectively. Calcium protected factor VII completely from the supernatant mediated inactivation. Cathepsin G and HLE purified from PMN each destroyed the coagulant activity of factor VII when added to a non-plasma system. There were, however, no effect on factor VII activity when cathepsin G was added to plasma. Polyacrylamide gel electrophoresis in the presence of SDS indicated that HLE and cathepsin G cleaved the zymogen in the same manner, producing (a) peptide(s) of low molecular mass and a single large product of 48 kDa. Preincubation of factor VII with calcium ions inhibited the proteolytic action of HLE and cathepsin G. It is suggested that HLE and cathepsin G from activated granulocytes may be partly responsible for the loss in factor VII activity that is observed during sepsis.

• References

• 1 Hagen FS, Gray CL, O’Hara P, Grant F, Saari GC, Woodbury RG, Hart CE, Inslay M, Kisiel W, Kurachi K, Davie EW. Characterization of a cDNA coding for human factor VII. Proc Natl Acad Sci USA 1986; 83: 2412-2416
  CrossrefPubMedGoogle Scholar
• 2 Fair DS. Quantitation of factor VII in the plasma of normal and warfarin-treated individuals by radioimmunoassay. Blood 1983; 62: 784-791
  PubMedGoogle Scholar
• 3 Zur M, Radcliffe RD, Oberdick J, Nemerson Y. The dual role of factor VII in blood coagulation. J Biol Chem 1982; 257: 5623-5631
  PubMedGoogle Scholar
• 4 Zur M, Nemerson Y. The esterase activity of coagulation factor VII: evidence for intrinsic activity of the zymogen. J Biol Chem 1978; 253: 2203-2209
  PubMedGoogle Scholar
• 5 Østerud B. Factor VII and haemostasis. Blood Coagul Fibrinol 1990; 1: 175-182
  PubMedGoogle Scholar
• 6 Laake K, Østerud B. Activation of purified plasma factor VII by human plasmin, plasma kallikrein and activated components of the human intrinsic blood coagulation system. Thromb Res 1974; 5: 759-772
  CrossrefPubMedGoogle Scholar
• 7 Seligsohn U, Østerud U, Brown SF, Griffin JH, Rapaport SI. Activation of human factor VII in plasma and in purified systems. Roles of activated factor IX, kallikrein, and activated factor XII. J Clin Invest 1979; 64: 1056-1065
  CrossrefPubMedGoogle Scholar
• 8 Radcliffe R, Nemerson Y. Mechanism of activation of bovine factor VII. J Biol Chem 1976; 251: 4797-4802
  PubMedGoogle Scholar
• 9 Radcliffe R, Nemerson Y. Activation and control of factor VII by activated factor X and thrombin. J Biol Chem 1975; 250: 388-395
  PubMedGoogle Scholar
• 10 Radcliffe R, Bagdasarian A, Colman R, Nemerson Y. Activation of bovine factor VII by Hageman factor fragments. Blood 1977; 50: 611-617
  PubMedGoogle Scholar
• 11 Schmidt W, Egbring R, Havemann K. Effect of elastase-like and chymotrypsin-like neutral proteases from human granulocytes on isolated clotting factors. Thromb Res 1975; 6: 315-326
  CrossrefPubMedGoogle Scholar
• 12 Turkington PT, Cathepsin G. a regulator of human vitamin K dependent clotting factors and inhibitors. Thromb Res 1992; 67: 147-155
  CrossrefPubMedGoogle Scholar
• 13 Turkington PT, Blumson NL, Elmore DT. The degradation of bovine and human prothrombin by human polymorphonuclear leukocyte cathepsin G. Thromb Res 1986; 44: 339-345
  CrossrefPubMedGoogle Scholar
• 14 Turkington PT. Degradation of human factor X by human polymorphonuclear leukocyte cathepsin G and elastase. Haemostasis 1991; 21: 111-116
  PubMedGoogle Scholar
• 15 Turkington PT. Cathepsin G can produce a gla-domainless protein C in vitro. Thromb Res 1991; 63: 399-406
  CrossrefPubMedGoogle Scholar
• 16 Damiano W, Kucich U, Murer E, Laudenslager N, Weinbaum G. Ultrastructural quantitation of peroxidase- and elastase-containing granules in human neutrophils. Am J Pathol 1988; 131: 234-245
  PubMedGoogle Scholar
• 17 Campbell EJ, Silveman EK, Campbell MA. Elastase and cathepsin G of human monocytes. Quantification of cellular content, release in response to Stimuli and heterogeneity in elastase-mediated proteolytic activity. J Immunol 1989; 143: 2961-2968
  PubMedGoogle Scholar
• 18 Campbell EJ, Senior RM, Welgus HG. Extracellular matrix injury during lung inflammation. Chest 1987; 92: 161-167
  CrossrefPubMedGoogle Scholar
• 19 Weitz JI, Huang AJ, Landman L, Nicholson S, Silverstein SC. Elastase mediated fibrinogenolysis by chemoattractant stimulated neutrophils occurs in the presence of physiologic concentrations of antiproteases. J Exp Med 1987; 166: 1836-1849
  CrossrefPubMedGoogle Scholar
• 20 Campbell EJ, Campbell MA. Pericellular proteolysis by neutrophils in the presence of proteinase inhibitors: effects of substrate opsonization. J Cell Biol 1988; 106: 667-676
  CrossrefPubMedGoogle Scholar
• 21 Powers JC, Grupton BF, Harley AD, Nishino N, Whitley RJ. Specificity of pancreatic elastase, human leukocyte elastase and cathepsin G. Inhibition with peptide chloromethyl ketones. Biochem Biophys Acta 1977; 485: 156-166
  CrossrefPubMedGoogle Scholar
• 22 Rao LVM, Bajaj SP. Purification of human factor VII utilizing o-(diethylaminoethyl)-sephadex and sulfopropyl-sephadex chromatography. Anal Biochem 1984; 136: 357-361
  CrossrefPubMedGoogle Scholar
• 23 Thim L, Bjørn S, Christensen M, Nicolaisen EM, Lund-Hansen T, Pedersen AH, Hedner U. Amino acid sequence and posttranslational modifications of human factor VII a from plasma and transfected baby hamster kidney cells. Biochemistry 1988; 27: 7785-7793
  CrossrefPubMedGoogle Scholar
• 24 Selak MA, Chignard M, Smith JB. Cathepsin G is a strong platelet agonist released by neutrophils. Biochem J 1988; 251: 293-299
  CrossrefPubMedGoogle Scholar
• 25 Boyum A. Isolation of lymphocytes, granulocytes and macrophages. Scand J Immunol 1976; (Suppl. 05) 9-15
  PubMedGoogle Scholar
• 26 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
  CrossrefPubMedGoogle Scholar
• 27 Hall CA, Rapaport SI, Ames SB, De Grot JA. A clinical and family study of hereditary proconvertin (factor VII) deficiency. Am J Med 1964; 37: 172-181
  CrossrefPubMedGoogle Scholar
• 28 Broze Jr GJ. Binding of human factor VII and VIIa to monocytes. J Clin Invest 1982; 70: 526-535
  CrossrefPubMedGoogle Scholar
• 29 Østerud B, Bjørklid E. Human factor VII associated with endotoxin stimulated monocytes in whole blood. Biochem Biophys Res Commun 1982; 108: 620-626
  CrossrefPubMedGoogle Scholar
• 30 Mc Gehee WG, Rapaport SI, Hjort PF. Intravascular coagulation in fulminant meningococcemia. Ann Intern Med 1967; 67: 250-260
  CrossrefPubMedGoogle Scholar