Identification of proteinase 3 as the major caseinolytic activity in acute human wound fluid.
Wound fluid contains several proteinases that are important in the repair process. In this study, we analyzed caseinolytic activity in wound fluid obtained from acute (burn) wounds. Caseinolytic activity in wound fluid increased markedly 2 d after injury and appeared on casein zymographs as a series of bands or a smear ranging from 30 to 100 kDa. Most of the enzyme activity was inhibited by the synthetic human neutrophil elastase inhibitor MDL 27,367 but not by the naturally occurring inhibitor of elastase, human secretory leukoproteinase inhibitor. Fractionation of wound fluid indicated that a single enzyme accounted for approximately 80% of the caseinolytic activity. This enzyme degraded the elastase substrate methoxysuccinyl-ala-ala-pro-val-p-nitroanilide at a slow rate. The above findings suggested that the enzyme responsible for caseinolytic activity might be proteinase 3, an elastase-related enzyme whose physiologic functions are poorly understood. Consistent with the above possibility, we found that monoclonal antibodies against proteinase 3 removed caseinolytic activity from wound fluid, and that purified proteinase 3 had a similar caseinolytic profile and inhibitor sensitivity to burn fluid.
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Department of Cell Biology and Neuroscience, University of Texas Southwestern Medical Center, Dallas 75235, USA.
Young
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Grinnell
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D000911:Antibodies, Monoclonal; D002056:Burns / Q000378:metabolism; D002364:Caseins / Q000378:metabolism*; D005122:Exudates and Transudates / Q000201:enzymology*; D006801:Humans; D007659:Ketones / Q000494:pharmacology; D053146:Myeloblastin; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors; D010455:Peptides / Q000494:pharmacology; D011233:Precipitin Tests; D012697:Serine Endopeptidases / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013997:Time Factors; D014945:Wound Healing / Q000502:physiology*
D000911:Antibodies, Monoclonal; D002364:Caseins; D007659:Ketones; C109772:MDL 27,367; D010455:Peptides; D015842:Serine Proteinase Inhibitors; D012697:Serine Endopeptidases; D010196:Pancreatic Elastase; D053146:Myeloblastin