Identification of neutrophil elastase as the proteinase in burn wound fluid responsible for degradation of fibronectin.
To identify proteinases responsible for fibronectin degradation in the wound environment we studied wound fluid obtained from burn patients. Immunoblotting experiments showed that extensive degradation of fibronectin had occurred in some burn wound fluid samples, in which case intact fibronectin molecules were undetectable, and the largest fibronectin fragment was 116 kDa. The 116-kDa fragment as well as a smaller 90-kDa fragment contained the fibronectin cell binding domain. These burn-fluid samples degraded freshly added fibronectin. Activity of the fibronectin-degrading enzyme was blocked by a broad-spectrum serine proteinase inhibitor or by specific neutrophil elastase inhibitors but not by metalloproteinase inhibitors or inhibitors of trypsin-like or chymotrypsin-like serine proteinases. Enzyme activity also was neutralized by antibodies against human neutrophil elastase. Incubation of fibronectin with burn wound fluid or purified human neutrophil elastase generated similar fibronectin-degradation products. Finally, direct assay of burn-wound-fluid samples with a synthetic elastase substrate showed a correlation between fluid-phase elastase activity and fibronectin degradation. Based on these findings, we conclude that burn-wound-fluid elastase is responsible for extensive fibronectin degradation. Acute elevation of elastase did not appear to hinder normal wound repair.
Department of Cell Biology and Neuroscience, University of Texas Southwestern Medical School, Dallas 75235-9039.
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Grinnell
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Zhu
D002056:Burns / Q000201:enzymology*; D002448:Cell Adhesion / Q000502:physiology; D010450:Endopeptidases / Q000032:analysis; D005122:Exudates and Transudates / Q000201:enzymology; D005353:Fibronectins / Q000097:blood / Q000378:metabolism*; D006801:Humans; D019272:Leukocyte Elastase; D010196:Pancreatic Elastase / Q000032:analysis*; D014945:Wound Healing / Q000502:physiology
D005353:Fibronectins; D010450:Endopeptidases; D010196:Pancreatic Elastase; D019272:Leukocyte Elastase