Elastase and neutral cathepsin production by human fibroblasts: effect of culture conditions on synthesis and secretion.
Fibroblasts from normal adult forearm skin and neonatal foreskin were cultured and examined for their ability to synthesize and secrete elastase and neutral cathepsin. All of the cultures examined produced detectable amounts of elastase using insoluble elastin as substrate. An enzyme was also found that hydrolyzed the synthetic elastin substrate, N-succinyl-(Ala)3-p-nitroanilide, but did not degrade insoluble elastin. In addition, activity against the synthetic cathepsin substrate N-benzoyl-DL-phenylalanine-naphthyl ester was found. Inhibitor profiles indicate that the elastin and N-succinyl-(Ala)3-p-nitroanilide degrading activities are due to metalloproteinases. Degradation of N-benzoyl-DL-phenylalanine-naphthyl ester can be inhibited by phenylmethylsulfonyl fluoride. These proteinases were usually found associated with the cell layer. Although activities of the measured proteinases were detected in all cultures, increased or decreased enzyme activities were not predictably related to passage number or length of serum starvation. Degree of confluence also affected proteinase activities. Separation of the dermal-epidermal junction can be produced by the injection of these proteinases into intact mouse skin.
D000328:Adult; D002403:Cathepsins / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D002478:Cells, Cultured; D004549:Elastin / Q000378:metabolism; D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D009842:Oligopeptides / Q000378:metabolism; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D010447:Peptide Hydrolases / Q000096:biosynthesis*; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000378:metabolism; D011480:Protease Inhibitors; D012867:Skin / Q000201:enzymology*
D009842:Oligopeptides; D011480:Protease Inhibitors; C049932:N-benzoylphenylalanine naphthyl ester; D010649:Phenylalanine; C015516:succinyl-trialanine-4-nitroanilide; D004549:Elastin; D002403:Cathepsins; D010447:Peptide Hydrolases; C023092:succinyltrialanine p-nitroanilide hydrolase; D010196:Pancreatic Elastase