title: Interaction of human leukocyte elastase with plasma fibronectin and its inhibition by soybean Bowman-Birk protease inhibitor.
abstract: Binding of human leukocyte elastase (HLE) to human plasma fibronectin (Fn) was determined by its competition with an oligopeptide chromogenic substrate. Kinetic curves of accumulation of the substrate hydrolysis product in the presence of Fn were specific for slow-binding one-step inhibitors of the enzyme. Values of rate constants for HLE association with Fn and for the dissociation of the complex were, respectively, 2.2.10(3) M(-1).sec(-1) and 1.4.10(-3) sec(-1) at pH 7.5 and 25 degreesC. The dissociation constant of the HLE--Fn complex determined independently by titration of the enzyme with the protein substrate and neglecting the hydrolysis was 3.9.10(-7) M. The resulting values were suggested to describe a high-affinity site in the Fn molecule which was subjected to the primary attack by HLE. The soybean Bowman--Birk protease inhibitor (BBI) efficiently inhibited the HLE-induced degradation of Fn under conditions of both inhibitor preincubation with the protease and its addition into the reaction mixture of the protease with Fn.
authors.affiliation: School of Chemistry, Lomonosov Moscow State University, Moscow, 119899, Russia. nilar@enzyme.chem.msu.su.
authors.forename: N I
authors.initials: NI
authors.lastname: Larionova
authors.forename: N G
authors.initials: NG
authors.lastname: Balabushevitch
authors.forename: P A
authors.initials: PA
authors.lastname: Zhatikov
mesh_terms: D001665:Binding Sites; D005353:Fibronectins / Q000097:blood*; D006801:Humans; D006868:Hydrolysis; D066298:In Vitro Techniques; D007700:Kinetics; D019272:Leukocyte Elastase / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D009842:Oligopeptides / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D013379:Substrate Specificity; D014358:Trypsin Inhibitor, Bowman-Birk Soybean / Q000494:pharmacology*
chemical_list: D005353:Fibronectins; D009842:Oligopeptides; D014358:Trypsin Inhibitor, Bowman-Birk Soybean; D019272:Leukocyte Elastase