Protein inhibitors of proteolytic enzymes

Means, Gary E.; Ryan, Dale S.; Feeney, Robert E.

doi:10.1021/ar50081a006

Cited by 23 publications

(10 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The spectrophotometric method was preferred since the substrate could be used in concentrations below the Km value, thus only slightly affecting the dissociation of the enzyme:inhibitor complex. These values are relatively higher than those obtained for other enzyme:inhibitor complexes (17,19,20). Indeed, significantly higher values for residual elastase I1 activity in the different enzyme:inhibitor complexes were obtained by the titrimetric method though the Km values for both substrates are similar.…”

Section: Inhibition O F Elastase Zz By Ovoinhibitor By Modified Ovoincontrasting

confidence: 51%

“…It has been shown that second order association kinetic constants for various chymotrypsin inhibitors fall within the range 0.022 -8.1 x lo6 min-' M-' (19). This was shown independently by electrophoretical and gel filtration studies in which preferential binding of chymotrypsin was observed, and to some extent, also by affinity chromatography.…”

Section: Discussionmentioning

confidence: 88%

See 1 more Smart Citation

Inhibition of Porcine Elastase Ii by Chicken Ovoinhibitor

Vered

Gertler

Burstein

1981

International Journal of Peptide and Protein Research

View full text Add to dashboard Cite

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

show abstract

Section: Inhibition O F Elastase Zz By Ovoinhibitor By Modified Ovoincontrasting

confidence: 51%

Section: Discussionmentioning

confidence: 88%

Inhibition of Porcine Elastase Ii by Chicken Ovoinhibitor

Vered

Gertler

Burstein

1981

International Journal of Peptide and Protein Research

View full text Add to dashboard Cite

show abstract

“…In the studies reported in this research, it has been possible to prepare modified inhibitors in which inhibitory activity against at least one of the enzymes remained largely intact, while a large decrease in inhibitory activity for one or more other enzymes has occurred. This has been previously accomplished by chemical or enzymatic modification of one of the binding sites (Means et al, 1974;Huber & Bode, 1978). Differential losses of inhibitory activity against trypsin, a-chymotrypsin and subtilisin upon treatment of turkey and penguin ovomucoid, and the conclusion that these losses were primarily due to decreases in association constants rather than to destruction of a binding site of the protein, appear to be further support for the importance of noncovalent associations in inhibitory mechanisms (Means et QL, 1974).…”

Section: Figurementioning

confidence: 99%

“…Protein inhibitors of proteolytic enzymes are well suited for the study of protein-protein interactions. Both the inhibitor-protease complex and the individual proteins may be studied using many physical and chemical techniques (Means et al 1974). Inhibitors have widely varying inhibitory activities against different enzymes.…”

mentioning

confidence: 99%

“…The acquisition of this number of sequences should eventually provide the information necessary for understanding differences not only in specificity but in strengths of associations between the inhibitors and the enzymes. An important aspect of the inhibitory activity of the protein inhibitors is not only their capacity to interact with these proteolytic enzymes but also their resistance to proteolysis (Means et al, 1974). Intimately related to this stabilization of the inhibitor molecules are, in most cases, certain critical disulfide bonds (Quast et al, 1978).…”

mentioning

confidence: 99%

See 1 more Smart Citation

INTRAMOLECULAR CROSS‐LINKING OF PROTEINS BY FORMATION OF LYSINOALANINE OR LANTHIONINE Modification of Disulfides in Ovomucoids

Walsh

Nashef

Feeney

1979

International Journal of Peptide and Protein Research

Self Cite

View full text Add to dashboard Cite

The disulfide bonds of ovomucoids were cleaved and new sulfur‐containing cross‐links were introduced by two separate chemical modification methods: (1) alkali treatment, and (2) cyanolysis. Alkali and cyanolytic treatments were used to cleave disulfide bonds and to introduce new synthetic nonreducible cross‐links consisting of residues of lysinoalanine (N‐(dl‐2‐amino‐2‐carboxyethyl)‐l‐lysine) and lanthionine (bis(2‐amino‐2‐carboxyethyl)sulfide). The two ovomucoids studied were turkey and penguin ovomucoids, which are “double‐headed” inhibitors of proteolytic enzymes with one inhibitory site for bovine trypsin and another for bovine α‐chymotrypsin or subtilisin. Trypsin does not compete with either α‐chymotrypsin or subtilisin, but the latter compete with one another, apparently for the same inhibitory site. The stability of inhibitory activities as a function of disulfide bond scission and the formation of new nondisulfide cross‐links was studied. In both methods of disulfide modification of turkey ovomucoid, inhibitory activity against trypsin was more stable with respect to the extent of modification than was inhibitory activity against chymotrypsin or subtilisin. With penguin ovomucoid, inhibitory activity against subtilisin was always more stable than activity against trypsin or chymotrypsin with both methods. With both ovomucoids it was thus possible to produce single‐headed inhibitors from the double‐headed inhibitors. The formation of the nonreducible cross‐links of lanthionine from cystines, and of lysinoalanines from cystines and lysines, with retention of a biochemical activity, suggests that such a procedure may have at least limited use as a cross‐linking method.

show abstract

Stability of Enzyme Inhibitors and Lectins in Foods and the Influence of Specific Binding Interactions

Zahnley

1984

Advances in Experimental Medicine and Biology

View full text Add to dashboard Cite

Protein inhibitors of proteolytic enzymes

Cited by 23 publications

References 45 publications

Inhibition of Porcine Elastase Ii by Chicken Ovoinhibitor

Inhibition of Porcine Elastase Ii by Chicken Ovoinhibitor

INTRAMOLECULAR CROSS‐LINKING OF PROTEINS BY FORMATION OF LYSINOALANINE OR LANTHIONINE Modification of Disulfides in Ovomucoids

Stability of Enzyme Inhibitors and Lectins in Foods and the Influence of Specific Binding Interactions

Contact Info

Product

Resources

About