On the presence of a metalloprotease in human skin fibroblasts that degrades the human skin elastic fiber system.
Succinyl-trialanine paranitroanilide, a specific synthetic substrate of elastases, was shown to be hydrolyzed by Triton X-100 extracts of human skin fibroblasts at near neutral pH. The neutral endopeptidase has been partially purified by ion exchange chromatography (DEAE Sephadex) and affinity chromatography using an AH-Sepharose (Ala)3 column. The enzyme has been purified 85-fold and appears to be a metalloprotease as shown by its inhibitory profile. In its partially purified form, the neutral endopeptidase was found inactive toward benzoyl arginine paranitroanilide, benzoyl tyrosine paranitroanilide, azocasein, type I collagen, and [3H]ligamentum nuchae-insoluble elastin. Structural glycoprotein microfibrils isolated from porcine aorta are extensively degraded by this neutral protease. It could also hydrolyze, but to a lesser extent, insoluble elastin purified from human aortas; it was, however, found inactive toward bovine ligamentum nuchae elastin. Its potentiality to degrade the human skin elastic fiber system (namely elastic fibers, oxytalan, and elaunin fibers) has been assessed by a morphometric analysis of the length of these fibers (on tissue sections appropriately stained to identify the components of the elastic fiber system) prior to and after enzyme action. Analysis of the data obtained by morphometry indicated that this neutral protease attacked rapidly both elaunin and oxytalan fibers of human dermis, but only slowly the mature elastic fibers.
D002478:Cells, Cultured; D002852:Chromatography, Ion Exchange; D004549:Elastin / Q000378:metabolism*; D010450:Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D005347:Fibroblasts / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D008666:Metalloendopeptidases; D010196:Pancreatic Elastase / Q000302:isolation & purification / Q000378:metabolism*; D012867:Skin / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism; D013379:Substrate Specificity
D004549:Elastin; D010450:Endopeptidases; D010196:Pancreatic Elastase; D008666:Metalloendopeptidases