Purification and characterization of the elastase-like enzyme of the bovine granulocyte.
The extracts of granules isolated from bovine granulocytes show elastase- and chymotrypsin-like activities, as detected with specific synthetic substrates. Extraction of these enzymes depends upon salt concentration. In the course of the present studies a 21-fold purification of the elastase-like enzyme was achieved on a (Ala)3-CH-Sepharose 4B gel. The molecular weight of the enzyme is 33 000, as determined by gel electrophoresis in the presence of sodium dodecyl sulfate. The elastase-like activity is inhibited by phenylmethylsulfonyl fluoride, soybean trypsin inhibitor, basic pancreatic inhibitor and by heparin at different rates. Elastatinal inhibits the enzyme competitively (Ki = 80 microM). The cytosol of bovine granulocytes contains a protein which strongly inhibits the elastase-like enzyme of the bovine granulocyte (Ki = 0.4 nM) as well as porcine pancreatic elastase (Ki = 11 nM).
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D003600:Cytosol / Q000378:metabolism; D006098:Granulocytes / Q000201:enzymology*; D006493:Heparin / Q000494:pharmacology; D009842:Oligopeptides / Q000494:pharmacology; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D011480:Protease Inhibitors / Q000378:metabolism; D013552:Swine; D014358:Trypsin Inhibitor, Bowman-Birk Soybean / Q000494:pharmacology; D014360:Trypsin Inhibitor, Kunitz Soybean / Q000494:pharmacology
D009842:Oligopeptides; D011480:Protease Inhibitors; D014358:Trypsin Inhibitor, Bowman-Birk Soybean; D010664:Phenylmethylsulfonyl Fluoride; D006493:Heparin; D014360:Trypsin Inhibitor, Kunitz Soybean; D010196:Pancreatic Elastase; C010718:elastatinal