Structural and Functional Dynamics of Lysosomal Cysteine Proteases with Particular Reference to Cathepsin B and Cathepsin H

2020 ◽  
pp. 391-424
Author(s):  
Sudhir K. Agarwal ◽  
Shalini Singh ◽  
Samir Sharma
Keyword(s):  
Cathepsin B ◽  
Cysteine Proteases ◽  
Functional Dynamics ◽  
Cathepsin H ◽  
Lysosomal Cysteine Proteases

scholarly journals In vitro effects of L-arginine on lysosomal cysteine proteases activity in isolated experiment and in the state of oxidative stress

2015 ◽  
Vol 96 (5) ◽  
pp. 876-882
Author(s):  
M A Fomina ◽  
A M Kudlaeva
Keyword(s):  
Oxidative Stress ◽  
Sucrose Solution ◽  
Membrane Damage ◽  
Cysteine Proteases ◽  
Lysosomal Membranes ◽  
In Vitro Incubation ◽  
Cathepsin H ◽  
Lysosomal Cysteine Proteases ◽  
Stimulation Of

Aim. Assessment of direct influence of arginine on lysosomal cysteine proteases activity in vitro, in isolation as well as the stimulation of oxidative stress. Methods. The study was conducted on the 72 female conventional mature Wistar rats 280-320 g divided into 6 series of 12 rats each. Lysosome slurries were isolated from the liver of intact animals with a subsequent in vitro incubation in a sucrose solution, in the presence of L-arginine, as well as in the presence of L-arginine accompanied by the stimulation of oxidative stress. Samples of control groups were exposed in vitro with the addition of isolate and oxidant, respectively. Each batch was reproduced three times, incubation was performed at 37 °C in a water bath for 1, 2 and 4 hours. The activity of cathepsins B, L and H was studied using spectrofluorimetric method in two fractions - intra- and extralysosomal. Acid phosphatase activity was used as the main marker of membrane labialization. Results. One hour Incubation with 5 mM arginine in vitro led to inhibition of the cathepsin H activity and lysosomal membrane damage, however, further increase in incubation time led to its stabilization. In vitro exposure to 5 mM H2O2 caused an increase in activity of cathepsines B and L and the drop in the cathepsin H activity without obvious changes in the distribution of enzymes between extra and intralysosomal fractions. In a state of oxidative stress 2-hour in vitro incubation with 5 mM arginine reduced the permeability of lysosomal membranes for cathepsines B, H and L; while 4-hour incubation led to the destabilization of lysosomal membranes. Conclusion. The direct effect of arginine at a concentration of 5 mM within the 1,2 and 4-hour time intervals leads to a distinct change as a lysosomal cysteine protease activity and stability of lysosomal membranes.

scholarly journals Effect of L-arginine and L-name on lysosomal cysteine proteases activity and lysosomal membranes permeability in rat aorta

2016 ◽  
Vol 97 (2) ◽  
pp. 250-255 ◽  
Author(s):  
A I Arapova ◽  
M A Fomina
Keyword(s):  
Acid Phosphatase ◽  
Cathepsin B ◽  
Cysteine Proteases ◽  
Total Activity ◽  
Rat Aorta ◽  
Lysosomal Fraction ◽  
Lysosomal Membranes ◽  
Cathepsin H ◽  
Male Wistar Rats ◽  
Combined Drugs

Aim. To study the effect of L-arginine and its analogue N-nitro-L-arginine methyl ester (L-NAME) alone and in combination on lysosomal cysteine proteolysis and lysosomal membranes state in rat aorta.Methods. The study was performed on male Wistar rats kept under standard vivarium conditions and divided into three control and three experimental groups of 8 animals each. The experimental samples included groups with L-arginine and/or L-NAME administration. The indicators were studied in the rat aorta homogenate precipitating and non precipitating fractions. Acid phosphatase activity was determined by a standardized method of «end point», the cathepsins B, L and H activity was studied by spectrofluorimetric method.Results. When simulating the changes of nitric oxide synthesis level using L-arginine, the increase of the total cathepsins activity was detected, acid phosphatase lability coefficient was reduced, what is characterized by general lysosomal membranes stabilization. L-NAME group, in contrast, is characterized by a decrease in the cathepsin B and H activity indicators, differences from arginine group were observed in the cathepsin H in lysosomal and general fractions, lysosomal membrane is labile. Combined drugs administration reduces the total cathepsins activity, while there is an increase of the acid phosphatase total activity, all indicators suggest lysosomal membranes labilization.Conclusion. L-arginine at a dose of 500 mg/kg causes increase in the total cathepsins B, L and H activity in rat aorta due to lysosomal fraction; L-arginine action leads to lysosomal membranes stabilization; L-NAME group in cathepsin H shows a decrease in the cathepsins secretion level with decreased total activity due to both factions; combined administration of arginine + L-NAME group in cathepsin B is characterized by an increase in secretion due to lysosomes membrane labilization.

Partial protease activity characterization of squid (Todaropsis eblanae) mantle / Caracterización parcial de la actividad proteolítica del manto de pota (Todaropsis eblanae)

1999 ◽  
Vol 5 (5) ◽  
pp. 391-396 ◽  
Author(s):  
M.G. Ayensa ◽  
H. An ◽  
M.C. Gómez-Guillén ◽  
P. Montero ◽  
A.J. Borderías
Keyword(s):  
Cathepsin B ◽  
Protease Activity ◽  
Cysteine Proteases ◽  
Acidic Ph ◽  
Alkaline Proteases ◽  
Alkaline Ph ◽  
Serine Protease Activity ◽  
Lysosomal Cysteine Proteases ◽  
Ph Range

Proteolytic activity in mantle of Todaropsis eblanae was maximum at 40 and 65 °C. Several peaks of activity were detected over the pH range studied (1.5-9.5), indicating the presence of acidic, neutral and alkaline proteases, depending on the temperature. The substantial enzymic inhibition at acidic pH by the inhibitor trans-epoxysuccinyl-L-leucylamine-4-guanidine butane (E-64) revealed the pre dominance of lysosomal cysteine proteases (cathepsins) which showed higher activity at 65 °C than at 40 °C. At 65 °C and pH 5.5 metallo-proteases were also detected by the inhibition with phenanthroline. Serine protease activity predominated at neutral pH (higher at 40 °C than at 65 °C), and cysteine proteases were detected at alkaline pH. There was evidence of cathepsin B and L activity at 65 °C and to a lesser degree at 40 °C.

Down-regulation of human extracellular cysteine protease inhibitors by the secreted staphylococcal cysteine proteases, staphopain A and B

2007 ◽  
Vol 388 (4) ◽  
pp. 437-446 ◽  
Author(s):  
Bjarne Vincents ◽  
Patrik Önnerfjord ◽  
Milosz Gruca ◽  
Jan Potempa ◽  
Magnus Abrahamson
Keyword(s):  
Cystatin C ◽  
Cathepsin B ◽  
Time Course ◽  
Cysteine Proteases ◽  
Normal Activity ◽  
Therapeutic Agents ◽  
Cysteine Protease Inhibitors ◽  
Protein Substrates ◽  
Cathepsin H

Abstract Of seven human cystatins investigated, none inhibited the cysteine proteases staphopain A and B secreted by the human pathogen Staphylococcus aureus. Rather, the extracellular cystatins C, D and E/M were hydrolyzed by both staphopains. Based on MALDI-TOF time-course experiments, staphopain A cleavage of cystatin C and D should be physiologically relevant and occur upon S. aureus infection. Staphopain A hydrolyzed the Gly11 bond of cystatin C and the Ala10 bond of cystatin D with similar K m values of approximately 33 and 32 μM, respectively. Such N-terminal truncation of cystatin C caused >300-fold lower inhibition of papain, cathepsin B, L and K, whereas the cathepsin H activity was compromised by a factor of ca. 10. Similarly, truncation of cystatin D caused alleviated inhibition of all endogenous target enzymes investigated. The normal activity of the cystatins is thus down-regulated, indicating that the bacterial enzymes can cause disturbance of the host protease-inhibitor balance. To illustrate the in vivo consequences, a mixed cystatin C assay showed release of cathepsin B activity in the presence of staphopain A. Results presented for the specificity of staphopains when interacting with cystatins as natural protein substrates could aid in the development of therapeutic agents directed toward these proteolytic virulence factors.

Cimaterol reduces cathepsin activities but has no anabolic effect in cultured myotubes

1990 ◽  
Vol 259 (6) ◽  
pp. E822-E827 ◽  
Author(s):  
D. M. Bechet ◽  
A. Listrat ◽  
C. Deval ◽  
M. Ferrara ◽  
J. F. Quirke
Keyword(s):  
Cathepsin B ◽  
Direct Action ◽  
Cathepsin L ◽  
Beta Agonist ◽  
Maximum Effect ◽  
Detectable Effect ◽  
Beta Agonists ◽  
Cathepsin H ◽  
Cultured Myotubes ◽  
Beta Adrenergic Agonist

The effect of the beta-adrenergic agonist cimaterol on bovine and chicken primary myotubes was assessed. Cimaterol at 10-100 nM concentrations reduced cathepsin B benzyloxy-carbonyl-Arg-Arg-4-methyl-7-coumarylamide hydrolyzing activity, as well as benzyloxycarbonyl-Phe-Arg-4-methyl-7-coumarylamide hydrolysis, which is a substrate for both cathepsin B and cathepsin L. Maximum effect was observed after 6-16 h treatment. Cathepsin H Arg-4-methyl-7-coumarylamide hydrolyzing activity was low and not significantly affected by cimaterol treatment. Despite decreasing cathepsin activities, cimaterol also increased proteolysis rates but induced no detectable effect on protein synthesis rates. These observations suggest that beta-agonists, as a result of a direct action on muscle, can decrease cathepsin activities but that beta-agonist-induced muscle hypertrophy may not be due to a direct effect on muscle cells.

scholarly journals Sodium salicylate activates cathepsin B but not cathepsin H from rat spleen

1984 ◽  
Vol 36 ◽  
pp. 49
Author(s):  
Kenji Yamamoto ◽  
Masahiro Tatsumi ◽  
Mitsue Takeda ◽  
Hisako Yamamoto ◽  
Osamu Kamata ◽  
...  
Keyword(s):  
Cathepsin B ◽  
Sodium Salicylate ◽  
Cathepsin H

Cathepsin B, Cathepsin H, Cathepsin X and Cystatin C in Sera of Patients with Early-Stage and Inflammatory Breast Cancer

2008 ◽  
Vol 23 (3) ◽  
pp. 161-168 ◽  
Author(s):  
J. Decock ◽  
N. Obermajer ◽  
S. Vozelj ◽  
W. Hendrickx ◽  
R. Paridaens ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cystatin C ◽  
Inflammatory Breast Cancer ◽  
Tumor Size ◽  
Cathepsin B ◽  
Early Stage ◽  
Menopausal Status ◽  
Preoperative Serum ◽  
Cathepsin H ◽  
Cathepsin X

Numerous studies have linked cathepsins and their inhibitor cystatin C to tumor invasion and metastasis. We examined whether cathepsin B, cathepsin H, cathepsin X and cystatin C could be detected in sera from women with early-stage or inflammatory breast cancer and whether they correlated with clinicopathological characteristics. Preoperative serum was obtained from 176 patients with early-stage breast cancer (tumor size <5 cm, negative lymph nodes) and 31 patients with inflammatory breast cancer. Cathepsin and cystatin C levels were measured by ELISA. The patient and tumor characteristics under study were age at diagnosis, menopausal status, tumor size, tumor grade, and steroid hormone receptor status. Serum cathepsin B levels were significantly lower in patients with poorly differentiated tumors. High cystatin C levels were associated with tumor size, postmenopausal status and patient age. Interestingly, significantly lower levels of cathepsin X and H were found in patients with inflammatory breast cancer, a trend also observed for cathepsin B and cystatin C. In conclusion, our results show a limited association of cathepsins B, H, X and cystatin C with established prognostic parameters. These data are promising and encourage future analysis of the clinical outcome of our patients in order to examine the potential prognostic value of these biomarkers. Further, this study indicates a role for cathepsin X and H in inflammatory breast cancer.

Cleavage of desmin by cysteine proteases: Calpains and cathepsin B

Meat Science ◽  
2004 ◽  
Vol 68 (3) ◽  
pp. 447-456 ◽  
Author(s):  
Caroline Pascale Baron ◽  
Susanne Jacobsen ◽  
Peter Patrick Purslow
Keyword(s):  
Cathepsin B ◽  
Cysteine Proteases

Lysosomal cysteine proteases (cathepsins): promising drug targets

2003 ◽  
Vol 59 (2) ◽  
pp. 203-213 ◽  
Author(s):  
Dušan Turk ◽  
Gregor Gunčar
Keyword(s):  
Drug Targets ◽  
Cysteine Proteases ◽  
Lysosomal Cysteine Proteases

OnBlastocystissecreted cysteine proteases: a legumain-activated cathepsin B increases paracellular permeability of intestinal Caco-2 cell monolayers

Parasitology ◽  
2016 ◽  
Vol 143 (13) ◽  
pp. 1713-1722 ◽  
Author(s):  
C. NOURRISSON ◽  
I. WAWRZYNIAK ◽  
A. CIAN ◽  
V. LIVRELLI ◽  
E. VISCOGLIOSI ◽  
...  
Keyword(s):  
Cathepsin B ◽  
Proteolytic Enzymes ◽  
Cell Monolayer ◽  
Cysteine Proteases ◽  
Cell Permeability ◽  
Whole Genome Sequence ◽  
Intestinal Cell ◽  
Pathogenic Potential ◽  
Cell Monolayers ◽  
Culture Supernatants

SUMMARYBlastocystisspp. pathogenic potential remains unclear as these anaerobic parasitic protozoa are frequently isolated from stools of both symptomatic and asymptomatic subjects.In silicoanalysis of the whole genome sequence ofBlastocystissubtype 7 revealed the presence of numerous proteolytic enzymes including cysteine proteases predicted to be secreted. To assess the potential impact of proteases on intestinal cells and gut function, we focused our study on two cysteine proteases, a legumain and a cathepsin B, which were previously identified inBlastocystissubtype 7 culture supernatants. Both cysteine proteases were produced as active recombinant proteins. Activation of the recombinant legumain was shown to be autocatalytic and triggered by acidic pH, whereas proteolytic activity of the recombinant cathepsin B was only recorded after co-incubation with the legumain. We then measured the diffusion of 4-kDa FITC-labelled dextran across Caco-2 cell monolayers following exposition to eitherBlastocystisculture supernatants or each recombinant protease. BothBlastocystisculture supernatants and recombinant activated cathepsin B induced an increase of Caco-2 cell monolayer permeability, and this effect was significantly inhibited by E-64, a specific cysteine protease inhibitor. Our results suggest that cathepsin B might play a role in pathogenesis ofBlastocystisby increasing intestinal cell permeability.

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