Different effects of hypochlorous acid on human neutrophil metalloproteinases: activation of collagenase and inactivation of collagenase and gelatinase.
Human neutrophils stimulated with phorbol 12-myristate 13-acetate (PMA) produce the reactive oxidant hypochlorous acid (HOCl) and release the matrix metalloproteinases collagenase and gelatinase from secretory granules. We have investigated the stoichiometry of activation and inactivation of the two metalloproteinases with HOCl. HOCl activated purified neutrophil procollagenase at ratios between 10 and 40 mol of HOCl/mol enzyme, but caused inactivation at higher ratios. Maximum activation was about the same as that achieved by p-aminophenyl-mercuric acetate. However, less than a third of the total collagenase released from PMA-stimulated neutrophils was activated by coreleased HOCl and most of the activity was destroyed after 1 h of stimulation. These results indicate that the HOCl/enzyme ratio must fall within a narrow range for activation to occur. In contrast to collagenase, purified progelatinase underwent negligible activation (2.5 +/- 1.2%) at HOCl/enzyme molar ratios less than 30 and was destroyed at higher ratios. Likewise no active gelatinase could be detected in supernatant from PMA-stimulated cells and almost all of the proenzyme was destroyed by HOCl after 60 min stimulation. Our results illustrate that only collagenase can be activated by HOCl in vitro and that gelatinase is much more sensitive to inactivation. Since a precise HOCl/enzyme ratio is required for collagenase activation it is doubtful whether effective enzyme regulation by HOCl could occur in vivo where various HOCl scavengers are present.
Michaelis
J
J
Department of Pathology, Christchurch School of Medicine, New Zealand.
Vissers
M C
MC
Winterbourn
C C
CC
D004591:Electrophoresis, Polyacrylamide Gel; D018093:Gelatinases; D006801:Humans; D006997:Hypochlorous Acid / Q000494:pharmacology*; D015151:Immunoblotting; D007700:Kinetics; D003012:Microbial Collagenase / Q000037:antagonists & inhibitors / Q000097:blood* / Q000302:isolation & purification; D008970:Molecular Weight; D009504:Neutrophils / Q000201:enzymology*; D010434:Pepsin A / Q000037:antagonists & inhibitors / Q000097:blood* / Q000302:isolation & purification; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014357:Trypsin / Q000378:metabolism
D006997:Hypochlorous Acid; D014357:Trypsin; D010434:Pepsin A; D018093:Gelatinases; D003012:Microbial Collagenase; D013755:Tetradecanoylphorbol Acetate