Identification of neutral proteases in human neutrophil granules that degrade articular cartilage proteoglycan.
Human polymorphonuclear neutrophil (PMN) granule extract (25 mug of protein) released 60 percent of the available 35SO4 from labeled rabbit articular cartilage in 0.5 hour at neutral pH. N-acetyl-L-alanyl-L-alanyl-L-prolyl-L-alanine choloromethyl ketone (NAcAAPACK), a specific elastase inhibitor, was only minimally effective against whole granule extract, and N-alpha-tosyl-L-lysine chloromethyl ketone, which inhibits trypsin but not elastase, was completely ineffective. Preparative disc-gel electrophoresis of PMN granule extract revealed two separate regions with independent activity against 35SO4-labeled cartilage. One region contained elastases and when tested alone, was completely inhibited by NAcAAPACK. The other contained lysozyme and two esterases active against N-acetyl-L-phenylalanine-alpha-naphthol. Purified lysozyme proved inactive, suggesting that the chymotrypsin-like esterases were responsible for proteoglycan degradation by this region of the gel.
D000818:Animals; D002358:Cartilage, Articular / Q000378:metabolism*; D003594:Cytoplasmic Granules / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004950:Esterases / Q000302:isolation & purification; D006025:Glycosaminoglycans / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007659:Ketones / Q000494:pharmacology; D009504:Neutrophils / Q000201:enzymology*; D010196:Pancreatic Elastase / Q000302:isolation & purification; D010447:Peptide Hydrolases / Q000302:isolation & purification*; D011509:Proteoglycans / Q000378:metabolism*; D011817:Rabbits; D013462:Sulfur Radioisotopes; D014108:Tosylphenylalanyl Chloromethyl Ketone / Q000494:pharmacology
D004791:Enzyme Inhibitors; D006025:Glycosaminoglycans; D007659:Ketones; D011509:Proteoglycans; D013462:Sulfur Radioisotopes; D014108:Tosylphenylalanyl Chloromethyl Ketone; D004950:Esterases; D010447:Peptide Hydrolases; D010196:Pancreatic Elastase