title: Proteinase inhibitory activities of antileukoprotease are represented by its second COOH-terminal domain.
abstract: Antileukoprotease or secretory leukocyte proteinase inhibitor is a potent serine proteinase inhibitor produced by exocrine glands of the human body. This monomeric protein (107 amino acids) comprises two homologous domains. It is generally thought that Leu19-Arg20-Tyr21 in the NH2-terminal domain represent the trypsin inhibitory activity, whereas Leu72-Met73-Leu74 in the COOH-domain represent the chymotrypsin and elastase inhibitory activity. Besides Met73, antileukoprotease contains three additional methionine residues all located in the COOH-terminal domain. Treatment of antileukoprotease with different amounts of methionine-selective reagents such as myeloperoxidase in the presence of H2O2 and Cl-, or cis-platinumdiammine dichloride resulted in a dose-dependent inactivation of all inhibitory activities, suggesting that methionine residues are involved in these activities. By using specific synthetic substrates, it was observed that elastase is able to displace trypsin from the inhibitor molecule, indicating that the trypsin and elastase inhibitory sites are located close to each other or at the same site. Incubation of antileukoprotease or its recombinant COOH-terminal domain with an antileukoprotease-specific monoclonal antibody (MoAb15) resulted in a strong selective increase of the trypsin inhibitory activity. The results presented reveal strong evidence that the inhibitory activities of antileukoprotease against trypsin, chymotrypsin and elastase are represented by its COOH-terminal domain, and that methionine residues are involved in interactions with these proteinases.
authors.affiliation: Department of Pulmonology, University Hospital Leiden, The Netherlands.
authors.forename: J A
authors.initials: JA
authors.lastname: Kramps
authors.forename: C
authors.initials: C
authors.lastname: van Twisk
authors.forename: H
authors.initials: H
authors.lastname: Appelhans
authors.forename: B
authors.initials: B
authors.lastname: Meckelein
authors.forename: T
authors.initials: T
authors.lastname: Nikiforov
authors.forename: J H
authors.initials: JH
authors.lastname: Dijkman
mesh_terms: D000911:Antibodies, Monoclonal; D001980:Bronchi / Q000201:enzymology; D002918:Chymotrypsin / Q000037:antagonists & inhibitors; D002945:Cisplatin / Q000494:pharmacology; D006801:Humans; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors; D009195:Peroxidase; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; D011994:Recombinant Proteins / Q000378:metabolism; D015842:Serine Proteinase Inhibitors / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014361:Trypsin Inhibitors
chemical_list: D000911:Antibodies, Monoclonal; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; D011994:Recombinant Proteins; D015842:Serine Proteinase Inhibitors; D014361:Trypsin Inhibitors; D009195:Peroxidase; D002918:Chymotrypsin; D010196:Pancreatic Elastase; D002945:Cisplatin