Human tryptase fibrinogenolysis is optimal at acidic pH and generates anticoagulant fragments in the presence of the anti-tryptase monoclonal antibody B12.
Human tryptase is uniquely regulated by its association with heparin and resists inhibition by biological protease inhibitors. The effects of pH and B12, an IgG anti-tryptase mAb, on cleavage of the synthetic substrate tosyl-Gly-Pro-Lys-p-nitroanilide and of the biological substrate fibrinogen by tryptase were examined. Tosyl-Gly-Pro-Lys-pnitroanilide cleavage was optimal at neutral pH and was inhibited by the B12 mAb at acidic and neutral pH values. At pH 7.5, inhibition was reversible and noncompetitive. In contrast, the optimal pH for tryptase to cleave fibrinogen was acidic. B12 dramatically enhanced the rate and extent that tryptase cleaved all three fibrinogen subunits at pH 6.0 to 6.5, but inhibited these activities at neutral pH. Major fibrinogen cleavage fragments generated at acidic pH by the B12:tryptase complex were identical with those made by plasmin. Thus, at acid pH, tryptase alone destroyed the ability of fibrinogen to clot, while the B12:tryptase complex increased the rate of fibrinogenolysis and also generated the anticoagulant, fragment D. The acidic pH optimum for tryptase fibrinogenolysis may direct this activity to tissue sites of inflammation. A putative biological equivalent to B12 would limit tryptase fibrinogenolytic activity at sites of neutral pH, such as blood, but would augment activity at acidic sites.
Ren
S
S
Department of Internal Medicine, Virginia Commonwealth University, Richmond 23298, USA.
Lawson
A E
AE
Carr
M
M
Baumgarten
C M
CM
Schwartz
L B
LB
D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000378:metabolism / Q000494:pharmacology*; D001667:Binding, Competitive / Q000276:immunology; D002384:Catalysis; D053818:Chymases; D016264:Dextran Sulfate; D004355:Drug Stability; D005340:Fibrinogen / Q000276:immunology / Q000378:metabolism*; D005341:Fibrinolysin / Q000378:metabolism; D005342:Fibrinolysis / Q000276:immunology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006969:Hypersensitivity, Immediate / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009842:Oligopeptides / Q000276:immunology / Q000378:metabolism; D010446:Peptide Fragments / Q000276:immunology* / Q000378:metabolism* / Q000502:physiology; D012697:Serine Endopeptidases / Q000276:immunology / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000276:immunology / Q000494:pharmacology*; D012882:Skin Tests; D013379:Substrate Specificity; D053802:Tryptases
D000276:Adjuvants, Immunologic; D000911:Antibodies, Monoclonal; D009842:Oligopeptides; D010446:Peptide Fragments; D015842:Serine Proteinase Inhibitors; D005340:Fibrinogen; D016264:Dextran Sulfate; D012697:Serine Endopeptidases; C083294:chymase 2; D053818:Chymases; D053802:Tryptases; D005341:Fibrinolysin