Interactions of human mast cell tryptase with biological protease inhibitors.
Tryptase from human mast cells has been shown (in vitro) to catalyze the destruction of fibrinogen and high-molecular-weight kininogen as well as the activation of C3a and collagenase. Although large amounts of tryptase are released in tissues by degranulating mast cells and levels as high as 1000 ng/ml have been measured in the circulation following systemic anaphylaxis, no specific physiologic inhibitor has yet been found for the protease. The current work tests several more inhibitors for their effects on tryptase and examines any effect of tryptase on these inhibitors. First, antileukoprotease and low-molecular-weight elastase inhibitor from human lung and hirudin and antithrombin III had no effect on tryptase activity in vitro. Second, the possibility that tryptase, being insensitive to the effects of inhibitors, might instead destroy them was also considered. Tryptase failed to cleave and inactivate antileukoprotease, low-molecular-weight elastase inhibitor, alpha 1 protease inhibitor, alpha 2 macroglobulin, and antithrombin III. Third, based on the knowledge that tryptase stability is regulated by its interaction with heparin, antithrombin III was used as a model heparin-binding protein to demonstrate that a protein competitor for heparin-binding sites, presumably by displacement of tryptase, destabilizes this enzyme. Conversely, tryptase, in excess, blocked the binding of antithrombin III to heparin, thereby attenuating the heparin-mediated inhibition of thrombin by antithrombin III.
Alter
S C
SC
Department of Medicine, Medical College of Virginia, Richmond 23298.
Kramps
J A
JA
Janoff
A
A
Schwartz
L B
LB
D000990:Antithrombin III / Q000378:metabolism; D004795:Enzyme Stability; D006801:Humans; D007700:Kinetics; D008407:Mast Cells / Q000201:enzymology*; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000378:metabolism*; D011480:Protease Inhibitors / Q000378:metabolism* / Q000494:pharmacology; D000511:alpha-Macroglobulins / Q000378:metabolism
D011480:Protease Inhibitors; D000511:alpha-Macroglobulins; D000990:Antithrombin III; D010447:Peptide Hydrolases; C023073:tosylarginine methyl ester hydrolase