Hurpin Is a Selective Inhibitor of Lysosomal Cathepsin L and Protects Keratinocytes from Ultraviolet-Induced Apoptosis†

The low survival rate of cardiac stem cells (CSCs) in the infarcted myocardium hampers cell therapy for ischemic cardiomyopathy. MicroRNA-21 (miR-21) and one of its target proteins, PTEN, contribute to the survival and proliferation of many cell types, but their prosurvival effects in c-kit+CSC remain unclear. Thus, we hypothesized that miR-21 reduces hydrogen peroxide- (H2O2-) induced apoptosis in c-kit+CSC and estimated the contribution of PTEN/PI3K/Akt signaling to this oxidative circumstance. miR-21 mimics efficiently reduced H2O2-induced apoptosis in c-kit+CSC, as evidenced by the downregulation of the proapoptosis proteins caspase-3 and Bax and upregulation of the antiapoptotic Bcl-2. In addition, the gain of function of miR-21 in c-kit+CSC downregulated the protein level of PTEN although its mRNA level changed slightly; in the meantime, miR-21 overexpression also increased phospho-Akt (p-Akt). The antiapoptotic effects of miR-21 were comparable with Phen (bpV), the selective inhibitor of PTEN, while miR-21 inhibitor or PI3K’s inhibitor LY294002 efficiently attenuated the antiapoptotic effect of miR-21. Taken together, these results indicate that the anti-H2O2-induced apoptosis effect of miR-21 in c-kit+CSC is contributed by PTEN/PI3K/Akt signaling. miR-21 could be a potential molecule to facilitate the c-kit+CSC therapy in ischemic myocardium.

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Сhanges in subcellular distribution of lysosomal cysteine proteinases activity in parenchymatous organs of rats under the action of nitric oxide synthesis modulators

Research n Practical Medicine Journal ◽

10.17709/2409-2231-2018-5-3-3 ◽

2018 ◽

Vol 5 (3) ◽

pp. 28-39

Author(s):

M. A. Fomina ◽

A. A. Terent'ev

Keyword(s):

Nitric Oxide ◽

Subcellular Distribution ◽

Cathepsin L ◽

Kidney Tissue ◽

Selective Inhibitor ◽

No Synthase ◽

Nitric Oxide Synthesis ◽

Lysosomal Activity ◽

Lysosomal Fraction ◽

Cathepsin H

Aim. To study the eﬀect of non-selective inhibitor of NO-synthase N-nitro-L-arginine methyl ester (L-NAME) and substrate of nitric oxide synthesis L-arginine on the activity of cathepsins B, L, H and its subcellular distribution in liver, kidney and lung tissues.Materials and methods. The object of study – male rats Wistar line, the material was the cytoplasmic and lysosomal fraction of homogenates of liver, kidney, lung tissues. A non-selective inhibitor of inducible NO-synthase N-nitro-L-arginine methyl ester (L-NAME) was applied at a dose of 25 mg/kg, the substrate of NO synthesis L-arginine – at a dose of 500 mg/kg. Activity of cathepsins B, L, H was defined separately in the cytoplasmic and lysosomal fractions by spectrofluorometry quantitative determination of the specific substrate cleavage product 7-amido-4-methylcoumarin.Results. Suppression of nitric oxide synthesis by non-selective inhibitor of NO-synthase L-NAME (25 mg/kg, 7 days) in the kidney tissue leads to a decrease in the activity of cathepsins В, L, H in lysosomal fraction with a parallel increase in non-lysosomal activity of cathepsin L, in the liver tissue leads to an increase in lysosomal activity of cathepsin H and a decrease in non-lysosomal activity of cathepsin L. The substrate of nitric oxide synthesis L-arginine (500 mg/kg, 10 days) only causes increased activity of cathepsin L in non-lysosomal fraction of liver tissue, leads to increased lysosomal activity of cathepsin H in kidney tissue, the lung tissue shows a significant increase in the activity of the all studied cathepsins in non-lysosomal fraction, accompanied by an increase in lysosomal activity of cathepsins B and H. The revealed changes are associated with the signs of changes in the ratio of pro-enzyme and active forms of cathepsins.Conclusion. The eﬀects of non-selective inhibitor and substrate of nitric oxide synthesis on the total activity of cathepsins B, L and H in parenchymatous organs and its subcellular distribution are tissue-specific and multidirectional in some cases and are accompanied by signs of changes in the ratio of pro-enzyme and enzymatically active forms mainly due to an increase of pro-enzyme forms.

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Calpain-Mediated Bid Cleavage and Calpain-Independent Bak Modulation: Two Separate Pathways in Cisplatin-Induced Apoptosis

Molecular and Cellular Biology ◽

10.1128/mcb.22.9.3003-3013.2002 ◽

2002 ◽

Vol 22 (9) ◽

pp. 3003-3013 ◽

Cited By ~ 163

Author(s):

Aleksandra Mandic ◽

Kristina Viktorsson ◽

Linda Strandberg ◽

Thomas Heiden ◽

Johan Hansson ◽

...

Keyword(s):

Cathepsin L ◽

Inhibitory Effect ◽

Dominant Negative ◽

Dominant Negative Mutant ◽

Drug Induced ◽

Calpain Activation ◽

Isolated Mitochondria ◽

Induced Apoptosis ◽

Calpain Inhibitors

ABSTRACT Calpain is a ubiquitous protease with potential involvement in apoptosis. We report that in human melanoma cells, cisplatin-induced calpain activation occurs early in apoptosis. Calpain activation and subsequent apoptosis were inhibited by calpeptin and PD150606, two calpain inhibitors with different modes of action. Furthermore, cisplatin induced cleavage of the BH3-only protein Bid, yielding a 14-kDa fragment similar to proapoptotic, caspase-cleaved Bid. However, Bid cleavage was inhibited by inhibitors of calpain, but not by inhibitors of caspases or of cathepsin L. Recombinant Bid was cleaved in vitro by both recombinant calpain and by lysates of cisplatin-treated cells. Cleavage was calpeptin sensitive, and the cleavage site was mapped between Gly70 and Arg71. Calpain-cleaved Bid induced cytochrome c release from isolated mitochondria. While calpeptin did not affect cisplatin-induced modulation of Bak to its proapoptotic conformation, a dominant-negative mutant of MEKK1 (dnMEKK) inhibited Bak modulation. dnMEKK did not, however, block Bid cleavage. The combination of dnMEKK and calpeptin had an additive inhibitory effect on apoptosis. In summary, calpain-mediated Bid cleavage is important in drug-induced apoptosis, and cisplatin induces at least two separate apoptotic signaling pathways resulting in Bid cleavage and Bak modulation, respectively.

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Azadirachtin-induced apoptosis involves lysosomal membrane permeabilization and cathepsin L release in Spodoptera frugiperda Sf9 cells

The International Journal of Biochemistry & Cell Biology ◽

10.1016/j.biocel.2015.03.018 ◽

2015 ◽

Vol 64 ◽

pp. 126-135 ◽

Cited By ~ 26

Author(s):

Zheng Wang ◽

Xingan Cheng ◽

Qianqian Meng ◽

Peidan Wang ◽

Benshui Shu ◽

...

Keyword(s):

Spodoptera Frugiperda ◽

Cathepsin L ◽

Membrane Permeabilization ◽

Lysosomal Membrane ◽

Sf9 Cells ◽

Lysosomal Membrane Permeabilization ◽

Induced Apoptosis

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The propeptide of Fasciola hepatica cathepsin L is a potent and selective inhibitor of the mature enzyme

Molecular and Biochemical Parasitology ◽

10.1016/s0166-6851(98)00164-9 ◽

1999 ◽

Vol 98 (2) ◽

pp. 271-277 ◽

Cited By ~ 26

Author(s):

Leda Roche ◽

Jose Tort ◽

JohnP. Dalton

Keyword(s):

Fasciola Hepatica ◽

Cathepsin L ◽

Selective Inhibitor ◽

Mature Enzyme

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Abstract 1337: MEK1:MAPK1/2 targeting attenuates pro-survival autophagy and enhances antiestrogen-induced apoptosis in breast cancer cells via a cathepsin L-dependent mechanism

10.1158/1538-7445.am2018-1337 ◽

2018 ◽

Author(s):

Annie Liu ◽

Carol Joseph ◽

Jesse Wayson ◽

Timothy Summers ◽

Haifeng Cai ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Breast Cancer Cells ◽

Cathepsin L ◽

Induced Apoptosis ◽

Dependent Mechanism

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Cathepsin L is involved in cathepsin D processing and regulation of apoptosis in A549 human lung epithelial cells

Biological Chemistry ◽

10.1515/bc.2004.082 ◽

2004 ◽

Vol 385 (7) ◽

Cited By ~ 46

Author(s):

A. Wille ◽

A. Gerber ◽

A. Heimburg ◽

A. Reisenauer ◽

C. Peters ◽

...

Keyword(s):

Epithelial Cells ◽

Cathepsin D ◽

Human Lung ◽

Cathepsin L ◽

A549 Cells ◽

Lung Epithelial Cells ◽

Single Chain ◽

Lung Epithelial ◽

Induced Apoptosis ◽

Human Lung Epithelial Cells

AbstractCathepsins are implicated in a multitude of physiological and pathophysiological processes. The aim of the present study was to investigate the function of cathepsin L (catL) in the proteolytic network of human lung epithelial cells and its role in the regulation of apoptosis. We found that catL-deficient A549 cells as well as lung tissue extracts of catL[-/-] mice express increased amounts of single-chain cathepsin D (catD). Degradation experiments indicate that catL specifically degrades the singlechain isoform of catD. Furthermore, we found that catLdeficient cells showed increased sensitivity to apoptosis. Finally, we demonstrate that the inhibition of catD activity by pepstatin A decreased the number of apoptotic cells in catLdeficient A549 cells after anti-Fas treatment. In conclusion, catL is involved in catD processing and the accumulation of catD isoforms in catL-deficient cells is associated with increased rates of spontaneous and anti-Fas-induced apoptosis.

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