Substrate specificity of the elastase and the chymotrypsin-like enzyme of the human granulocyte

Zimmerman, M.D.; Ashe, Bonnie M.

doi:10.1016/0005-2744(77)90337-0

Cited by 98 publications

(53 citation statements)

References 17 publications

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“…In contrast, porcine pancreas elastase prefers Ala residues while cathepsin G (a chymotrypsin-like enzyme) and chymotrypsin have a preference for Phe residues in the P1 position (32,38,39). Macrophage elastase, a metalloenzyme containing a coordinated zinc atom in its active site, prefers to cleave peptide bonds to which Leu contributes the a-NH2-group (40).…”

Section: Discussionmentioning

confidence: 99%

Development of an assay for in vivo human neutrophil elastase activity. Increased elastase activity in patients with alpha 1-proteinase inhibitor deficiency.

Weitz¹,

Landman²,

Crowley³

et al. 1986

J. Clin. Invest.

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Leukocyte extracts contain enzymes that digest fibrinogen and release a fibrinopeptide A-containing fragment. This study was undertaken to identify the responsible proteinase and to characterize the fibrinopeptide A-containing fragment so that it could be used as an index of enzyme activity. Both the fibrinogenolytic activity and the release of the fibrinopeptide A-containing fragment mediated by the leukocyte extracts were shown to be due to human neutrophil elastase (HNE) by the following criteria: activity was completely blocked by a specific HNE inhibitor or by adsorbing HNE from the extracts with a monospecific antibody and reconstitution with purified HNE restored the ability to release the fibrinopeptide A-containing fragment. This fragment was not released by a variety of other proteinases or by HNE-inhibitor complexes indicating that, at least with respect to the enzymes tested, it is a specific product of HNE and its release requires the free enzyme. By separating the products of HNE digestion of fibrinogen using high performance liquid chromatography, identifying the immunoreactive fractions and subjecting them to amino acid analysis, the fragment was identified as Aal-21, indicating an HNE cleavage site at the Val(Aa21)-Glu(Aa22) bond. The mean plasma Aal-21 level was markedly higher in patients with a1-proteinase inhibitor deficiency as compared to healthy controls (0.2 nM vs. 7.9 nM; P < 0.0001), consistent with increased in vivo HNE activity in these individuals.

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Section: Discussionmentioning

confidence: 99%

Development of an assay for in vivo human neutrophil elastase activity. Increased elastase activity in patients with alpha 1-proteinase inhibitor deficiency.

Weitz¹,

Landman²,

Crowley³

et al. 1986

J. Clin. Invest.

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“…Complete HNE resistance was not obtained with the substitutions used, which is perhaps not surprising considering the general preference of HNE for P1 residues with a medium-sized aliphatic side chain (48). There was a good correlation, however, between the order of P1 residues preferred by HNE in small peptide substrates and chloromethylketone inhibitors (Ile = Val > Leu > Ala) (48)(49)(50)(51), and the substitutions conferring HNE resistance in C I inhibitor (Leu or Ala for Ile or Val). In addition, our results were also in agreement with computer modeling that has predicted that P 1-Ile would be a better substrate for HNE than Pl-Leu (48).…”

Section: Discussionmentioning

confidence: 92%

Recombinant C1 inhibitor P5/P3 variants display resistance to catalytic inactivation by stimulated neutrophils.

Eldering

Huijbregts²,

Nuijens³

et al. 1993

J. Clin. Invest.

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“…In this study, we succeeded in purifying Codium fragile proteases using a sensitive substrate and affinity chromatography technique. It has been reported that Suc-Ala-Ala-Pro-Phe-pNA, Ac-AlaAla-Pro-Ala-pNA and Suc-Ala-Ala-Pro-Arg-MCA are good substrates for chymotrypsin (DelMar et al 1979), elastase (Zimmerman and Ashe 1977) and trypsin (Graf etal. 1988).…”

Section: Discussionmentioning

confidence: 99%

Some properties of trypsin-like proteases extracted from the seaweedCodium fragile and their purification

Kadokami¹,

Yoshida

Mizusaki

et al. 1990

Mar. Biol.

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Abstract. Two enzymes were purified from the green algaCodiumfragile, collected in 1988 at Fukuoka, Japan, by batch-wise ion-exchange extraction, affinity chromatography and ion-exchange high-performance liquid chromatography (HPLC) using Boc-Ala-Ala-Pro-Arg-pNA as a substrate. The molecular weights of the enzymes were estimated as 38 000 and 39 000, using gel filtration HPLC. The enzymes had the same optimal pH range of 7 to 9 for both activities, and an exclusively hydrolyzed peptide bond on the carboxyl-terminal side of the Larginine of peptide p-nitroanilides. The ratios of the enzymatic activity for X-Arg-pNA to X-Lys-pNA were larger than 100. The enzymes exhibited 30 times higher activity toward Boc-Ala-Ala-Pro-Arg-pNA when compared with trypsin. The activities were strongly inhibited by diisopropyl phosphofluoridate (DFP), and partially inhibited by phenylmethylsulfonyl fluoride (PMSF), benzamidine, leupeptin and antipain. The isolated enzymes were presumed to be trypsin-like serine protease from their primary substrate specificities and inactivation behavior.

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Substrate specificity of the elastase and the chymotrypsin-like enzyme of the human granulocyte

Cited by 98 publications

References 17 publications

Development of an assay for in vivo human neutrophil elastase activity. Increased elastase activity in patients with alpha 1-proteinase inhibitor deficiency.

Development of an assay for in vivo human neutrophil elastase activity. Increased elastase activity in patients with alpha 1-proteinase inhibitor deficiency.

Recombinant C1 inhibitor P5/P3 variants display resistance to catalytic inactivation by stimulated neutrophils.

Some properties of trypsin-like proteases extracted from the seaweedCodium fragile and their purification

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