Cathepsin B-like cysteine proteinase activity in sputum and bronchoalveolar lavage samples: relationship to inflammatory cells and effects of corticosteroids and antibiotic treatment

1985 ◽  
Vol 68 (4) ◽  
pp. 469-474 ◽  
Author(s):  
David Burnett ◽  
Robert A. Stockley

1. Cathepsin B-like activity was measured in lung secretions by using the fluorimetric substrate benzyloxycarbonyl-l-arginine-l-arginine-4-methyl-7-coumarylamide (Z-Arg-Arg-MEC). The enzyme had a pH optimum of approximately 5.5 and had the characteristics of an alkaline-stable cysteine proteinase. 2. Enzyme activity in the sputum from ten subjects with chronic bronchitis was significantly reduced after 5 days’ treatment with prednisolone. 3. Seven patients with bronchiectasis were studied before and after 14 days’ treatment with amoxycillin. Cysteine proteinase activity was significantly reduced after 7 days’ therapy, in parallel with a change in sputum quality from purulent to mucoid. One week after cessation of treatment enzyme levels were again increased but were still significantly lower than pretreatment values. 4. Enzyme activity in 21 bronchoalveolar lavage specimens correlated significantly with neutrophil counts but not with macrophage counts. 5. Cysteine proteinase activity in lung secretions resembles that of cathepsin B but is alkaline-stable, suggesting it is a distinct enzyme. The levels of cysteine proteinase in lung secretions appear to be related to the presence of inflammation or infection.

Parasitology ◽  
1994 ◽  
Vol 109 (3) ◽  
pp. 357-365 ◽  
Author(s):  
V. M. Koritsas ◽  
H. J. Atkinson

SummarySensitive assays capable of detecting proteinases in single females of the phytoparasite Globodera pallida have been developed and used to define the proteinase activity of young adult females. Digestion of the large subunit of the plant protein Rubisco established a pH optimum for the proteinase activity at pH 5·7. The activity was inhibited by the cysteine proteinase inhibitors p-chloromercuribenzoic acid (PMBA) and p-chloromercurisulphonic acid (PMSA) and stimulated by both cysteine and dithiothreitol (DTT). It was moderately reduced by L-trans-epoxysuccinyl-leucylamido-(4- guanidino) butane (E64) but not by specific inhibitors of serine, aspartate or metallo-proteinases. The activity separated into 3 bands on a non-denaturing gel but only I proteinase of 62 kDa was recovered following a combination of anion-exchange chromatography and affinity chromatography using PMBA. The effect of inhibitors was similar to that reported previously for some of the cysteine proteinase activity recovered from Caenorhabditis elegans but is apparently not that for which the corresponding gene has been cloned in this nematode and Haemonchus contortus. The proteinase may have a major role in digestion of dietary protein and so offers an exciting target for future control of this important plant-parasitic nematode.


2003 ◽  
Vol 279 (7) ◽  
pp. 5081-5087 ◽  
Author(s):  
Yasuhisa Sakata ◽  
Kazuhiko Arima ◽  
Toshiro Takai ◽  
Wataru Sakurai ◽  
Kiyonari Masumoto ◽  
...  

2009 ◽  
Vol 122 (4) ◽  
pp. 306-309 ◽  
Author(s):  
Michelle A.R. Freitas ◽  
Helen C. Fernandes ◽  
Viviane C. Calixto ◽  
Almir S. Martins ◽  
Edward F. Silva ◽  
...  

1982 ◽  
Vol 205 (2) ◽  
pp. 295-302 ◽  
Author(s):  
A D Gounaris ◽  
E E Slater

Cysteine-proteinase activity was observed in homogenates of human-cadaver renal cortex. This activity co-purified with renin enzymic activity until separation by aminohexyl-Sepharose-pepstatin affinity chromatography. The cysteine proteinase was purified 1780-fold after the following successive chromatographic procedures: Sephadex G-75, DEAE-cellulose DE-52, and an organomercurial affinity resin. The proteinase activity was dependent upon activation by thiol-containing compounds such as dithiothreitol, as well as by EDTA, and was inhibited by the thiol-group-specific alkylating reagents iodoacetic acid and N-ethylmaleimide. DE-52 cellulose chromatography resolved the cysteine proteinase into two components. On the basis of molecular size (26 000 daltons), activity as a function of pH, stability as a function of pH, substrate specificity and thermal lability, the major component (95%) has been identified as cathepsin B. The DE-52 cellulose elution pattern of the minor component (5%) is suggestive of cathepsin H [Schwartz & Barrett (1980) Biochem. J. 191, 487-497] Enzymic activity was determined with synthetic substrates, in particular alpha-N-benzoyl-DL-arginine 2-naphthylamide (Bz-Arg-NNap), thus precluding the detection of cathepsin L [Kirschke, Langner, Wiederanders, Ansorge, Bohley & Broghammer (1976) Acta Biol. Med. Germ. 35, 285-299]. Inhibition by dimethyl sulphoxide was observed in the determination of Km = 7.0 +/- 0.4 mM for the substrate Bz-Arg-NNap, and care must therefore be taken in the preparation of substrate solutions.


Heliyon ◽  
2021 ◽  
Vol 7 (10) ◽  
pp. e08125
Author(s):  
F.A.F. Mansur ◽  
W. Luoga ◽  
J.M. Behnke ◽  
D.J. Buttle ◽  
I.R. Duce ◽  
...  

1997 ◽  
Author(s):  
A Eijan ◽  
A Casabe ◽  
L Puricelli ◽  
L Pasik ◽  
H Malagrino ◽  
...  

Author(s):  
Luca Salvati ◽  
Marco Mattu ◽  
Marco Colasanti ◽  
Aldo Scalone ◽  
Giorgio Venturini ◽  
...  

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