Activation of human neutrophil gelatinase by endogenous serine proteinases.
The role of serine proteinases and oxidants in the activation of gelatinase released from human neutrophils was investigated. Gelatinase was measured by its ability to degrade both gelatin and native glomerular basement-membrane type IV collagen. When fMet-Leu-Phe or phorbol 12-myristate 13-acetate was used to stimulate the neutrophils, no gelatinase activity was measured in the absence of a mercurial activator, indicating that the enzyme was released entirely in latent form. However, when fMet-Leu-Phe-stimulated cells were treated with cytochalasin B, 50-70% of the maximal gelatinase activity was released. Activation was blocked by the serine-proteinase inhibitor phenylmethanesulphonyl fluoride and a specific inhibitor of neutrophil elastase, but was not affected by an inhibitor of cathepsin G. Addition of catalase or azide to prevent oxidative reactions did not affect activation of gelatinase under any conditions of stimulation, indicating that oxidants were not involved in activation. Our results imply that oxidative activation of gelatinase does not occur readily. However, neutrophil serine proteinases, particularly elastase, provide an alternative and apparently more efficient mechanism of activation.
Vissers
M C
MC
Department of Pathology, University of Michigan Medical School, Ann Arbor 48109.
Winterbourn
C C
CC
D001485:Basement Membrane / Q000378:metabolism; D003094:Collagen / Q000378:metabolism; D003571:Cytochalasin B / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D005780:Gelatin / Q000378:metabolism; D018093:Gelatinases; D006801:Humans; D066298:In Vitro Techniques; D007678:Kidney Glomerulus / Q000378:metabolism; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology*; D010434:Pepsin A / Q000097:blood*; D012697:Serine Endopeptidases / Q000097:blood*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D003571:Cytochalasin B; D009240:N-Formylmethionine Leucyl-Phenylalanine; D005780:Gelatin; D003094:Collagen; D012697:Serine Endopeptidases; D010434:Pepsin A; D018093:Gelatinases; D013755:Tetradecanoylphorbol Acetate