title: The role of lysosomal elastase in the digestion of Escherichia coli proteins by human polymorphonuclear leukocytes: experiments with living leukocytes.
abstract: Human polymorphonuclear leukocyte (PMN) elastase has been implicated in various pathological conditions. However, its physiological role remains undefined. One possible function of this enzyme may be digestion of bacterial proteins after phagocytosis. To test this hypothesis, we prepared Escherichia coli labeled with [3H]arginine and treated these bacteria with a lipid-soluble, active-site-directed chloromethyl ketone inactivator of pancreatic and granulocyte elastases (carbobenzoxy-L-glycyl-L-leucyl-L-alanine chloromethyl ketone, dissolved in methanol). Control bacteria were treated with methanol alone. When E. coli pretreated with the inactivator were incubated with solutions of porcine pancreatic elastase or with PMN granule extract, release of trichloroacetic acid-soluble radioactivity was significantly lower than in the control E. coli. Similar results were obtained when treated and control E. coli were fed to viable human PMN. In contrast, release of trichloroacetic acid-soluble radioactivity from E. coli containing [3H]thymidine was not affected by pretreatment of bacteria with elastase inactivator before feeding them to PMN, suggesting that phagocytosis of E. coli had not been inhibited by the chloromethyl ketone. When treated and control bacteria were fed to PMN, no significant difference was observed in the activity of lysosomal beta-glucuronidase recovered from post-granule supernatant fractions of homogenized leukocytes, suggesting that lysosomal degranulation had not been suppressed by the inactivator. However, elastase activity of the same fractions was depressed if the leukocytes had phagocytized chloromethyl ketone-treated E. coli, suggesting that inhibition of PMN elastase had occurred. We conclude that PMN elastase participates in digestion of E. coli proteins by human PMN.
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D002918:Chymotrypsin; D004926:Escherichia coli; D008247:Lysosomes / Q000201:enzymology*; D000432:Methanol; D009504:Neutrophils / Q000201:enzymology*; D010196:Pancreatic Elastase / Q000502:physiology*; D010587:Phagocytosis
chemical_list: D001426:Bacterial Proteins; D002918:Chymotrypsin; D010196:Pancreatic Elastase; D000432:Methanol