Inhibition of Porcine Elastase Ii by Chicken Ovoinhibitor

Abstract

Interaction o f porcine elastase II with native and modified chicken egg-white ovoinhibitor was studied by determining the residual activity of the partially inhibited enzyme and by direct measurement of the stoichiometry of interaction using affinity chromatography, electrophoresis and gel filtration. It was found that the chymotrypsin binding site that is not modified by mild oxidation with N-chlorosuccinimide (Shechter et al., Biochemistry, (1977) 16,[992][993][994][995][996][997] is capable o f binding elastase II as well. The binding o f chymotrypsin and elastase II is mutually exclusive and the affinity for chymotrypsin is stronger. Binding of 2 mol trypsin as 1 rnol elastase I by ovoinhibitor does not interfere with the binding of elastase II. There is also an indication that the second binding site for chymotrypsin is capable o f forming a complex with an additional molecule o f elastase I . but the binding is so weak that it could be detected only by electrophoresis.Key words: chicken ovoinhibitor; inhibition; porcine elastase 11; protein-protein inter- action.Chicken ovoinhibitor, an egg-white glyco-exhibits chymotrypsin-like specificity toward protein with a molecular weight of 48000, hydrolysis of synthetic substrates and its is a multiheaded inhibitor of several mammalian amino terminal sequence strongly resembles serine pancreatic proteinases (1-3). It has been the B chain of bovine chymotrypsin B. Howdemonstrated by kinetic studies (1, 2, 4, 5 ) ever, unlike chymotrypsin, it hydrolyzes and by direct stoichiometric measurements elastin and is not inhibited by turkey ovo-(6) that it can simultaneously inhibit two mucoid (8). molecules of trypsin, two molecules of c h y m e ,We have reported that ovoinhibitor is trypsin and one molecule of elastase I. capable of inhibiting elastase I1 (9). The present Porcine elastase I1 is a recently described study was devoted t o further characterization pancreatic proteinase (7, 8). This enzyme of the interaction between these proteins and identification of the ovoinhibitor binding site(s) Abbrevhti ons: Ac, N-acetyl; Glut, glutaryl; Tos, that reacts with elastase 11. This was achieved N-p-toluenesulfonyl; OMe, methyl ester; OEt, ethyl by using both kinetic studies and measuring ester. the stoichiometry of interaction by affinity 169 0367-8377/81/070169-11 $02.00/0 o 1981 Munkspmd, Copenhagen

Egg white versus Salmonella Enteritidis! A harsh medium meets a resilient pathogen

“…Each domain contains an inhibitory site. Ovoinhibitor has been shown to be active against a large spectrum of serine proteases such as trypsin (Strechter et Berger, 1967;Galzie et al 1996), chymotrypsin and elastase (ScottTomimatsu et al, 1966;feeney, 1971;Gertier and Feinstein, 1971;Vered et al, 1987Vered et al, , 1981.…”

The Differential Expression and Snp Analysis of the Ovoinhibitor Gene in the Ovaries of Laying Duck Breeds (Anas Platyrhynchos)

“…Ovoinhibitor (OIH) is a Kazal-type serine inhibitor. It is the main protease inhibitor found in the chicken plasma, and known to inhibit the activity of trypsin, chymotrypsin, and elastase (Zahnley, 1975;Shechter et al 1977;Vered et al, 1981). OIH is a three-domain protein and shares its structure with the ovomucoid protein (Moore et al, 2004).…”

Digestion of elastin by porcine pancreatic elastase I and elastase II

“…However a few years later, it was reported by Ardelt (1 1 , 12) that an additional pancreatic endopeptidase with chymotrypsin-like specificityelastase I1 -is also capable of solubilizing elastin. We have further purified, characterized (13, 14) and partially sequenced (15) this enzyme. It was confirmed that in spite of the completely different specificity towards synthetic substrates, both enzymes hydrolyze and solubilize elastin.…”