Verification of a Putative Caspase Substrate

2014 ◽  
Vol 2014 (8) ◽  
pp. pdb.prot080317-pdb.prot080317 ◽  
Author(s):  
G. P. McStay ◽  
D. R. Green
Keyword(s):  
Caspase Substrate

scholarly journals Stress-Inducible Caspase Substrate TRB3 Promotes Nuclear Translocation of Procaspase-3

PLoS ONE ◽  
2012 ◽  
Vol 7 (8) ◽  
pp. e42721 ◽  
Author(s):  
Kouhei Shimizu ◽  
Shoukichi Takahama ◽  
Yaeta Endo ◽  
Tatsuya Sawasaki
Keyword(s):  
Nuclear Translocation ◽  
Caspase Substrate

scholarly journals F1Aα, a Death Receptor-binding Protein Homologous to theCaenorhabditis elegansSex-determining Protein, FEM-1, Is a Caspase Substrate That Mediates Apoptosis

1999 ◽  
Vol 274 (45) ◽  
pp. 32461-32468 ◽  
Author(s):  
Shing-Leng Chan ◽  
Kuan-Onn Tan ◽  
Li Zhang ◽  
Karen S. Y. Yee ◽  
Francesca Ronca ◽  
...  
Keyword(s):  
Receptor Binding ◽  
Binding Protein ◽  
Death Receptor ◽  
Caspase Substrate ◽  
Receptor Binding Protein

Sequential and rapid activation of select caspases during apoptosis of normal intestinal epithelial cells

1998 ◽  
Vol 274 (6) ◽  
pp. G1117-G1124 ◽  
Author(s):  
Johannes Grossmann ◽  
Susanne Mohr ◽  
Eduardo G. Lapetina ◽  
Claudio Fiocchi ◽  
Alan D. Levine
Keyword(s):  
Epithelial Cells ◽  
Intestinal Epithelial Cells ◽  
Cysteine Proteases ◽  
Intestinal Epithelial ◽  
Caspase Activation ◽  
High Turnover ◽  
Sequential Activation ◽  
Caspase 6 ◽  
Caspase Substrate ◽  
Induced Apoptosis

Detachment-induced cell death (DICD) is considered to be one of the means by which intestinal epithelial cells (IEC) die of apoptosis as they reach the lumen and are shed. Caspases, a family of cysteine proteases, play a central role in initiating, amplifying, and executing apoptosis; however, the pattern of caspase activation in response to distinct apoptotic stimuli remains unknown. We investigated the kinetics of caspase activation during DICD in freshly isolated human IEC. DNA fragmentation is observed 90 min after detachment and is preceded by the sequential activation of preformed members of the CPP32 family of caspases. Activation of caspase 6 and cleavage of the endogenous caspase substrate poly(ADP-ribose) polymerase (EC 2.4.2.30 ) are detected within 15 min of detachment, 30–45 min before caspase 3 activation. Caspase 1 and caspase 10 are present as proenzymes, yet they remain inactive in response to this trigger of apoptosis. Human IEC are primed to rapidly undergo detachment-induced apoptosis involving the selective and sequential activation of preformed caspases. This study may enhance our understanding of physiological events occurring as IEC are shed. Their rapid apoptotic response to detachment may facilitate the high turnover of cells and ensure homeostasis in the intestinal epithelium.

scholarly journals Development of a Cationic Amphiphilic Helical Peptidomimetic (B18L) As A Novel Anti-Cancer Drug Lead

Cancers ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 2448 ◽  
Author(s):  
Yuan Lyu ◽  
Steven Kopcho ◽  
Folnetti A. Alvarez ◽  
Bryson C. Okeoma ◽  
Chioma M. Okeoma
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Cancer Progression ◽  
Cancer Drug ◽  
Cancer Cell Death ◽  
Drug Lead ◽  
Anti Cancer ◽  
Cancer Agent ◽  
Caspase Substrate ◽  
Dynamics Simulations

BST-2 is a novel driver of cancer progression whose expression confers oncogenic properties to breast cancer cells. As such, targeting BST-2 in tumors may be an effective therapeutic approach against breast cancer. Here, we sought to develop potent cytotoxic anti-cancer agent using the second-generation BST-2-based anti-adhesion peptide, B18, as backbone. To this end, we designed a series of five B18-derived peptidomimetics. Among these, B18L, a cationic amphiphilic α-helical peptidomimetic, was selected as the drug lead because it displayed superior anti-cancer activity against both drug-resistant and drug-sensitive cancer cells, with minimal toxicity on normal cells. Probing mechanism of action using molecular dynamics simulations, biochemical and membrane biophysics studies, we observed that B18L binds BST-2 and possesses membranolytic characteristics. Furthermore, molecular biology studies show that B18L dysregulates cancer signaling pathways resulting in decreased Src and Erk1/2 phosphorylation, increased expression of pro-apoptotic Bcl2 proteins, caspase 3 cleavage products, as well as processing of the caspase substrate, poly (ADP-ribose) polymerase-1 (PARP-1), to the characteristic apoptotic fragment. These data indicate that through the coordinated regulation of membrane, mitochondrial and signaling events, B18L executes cancer cell death and thus has the potential to be developed into a potent and selective anti-cancer compound.

scholarly journals SVM-based prediction of caspase substrate cleavage sites

2006 ◽  
Vol 7 (Suppl 5) ◽  
pp. S14 ◽  
Author(s):  
Lawrence JK Wee ◽  
Tin Tan ◽  
Shoba Ranganathan
Keyword(s):  
Cleavage Sites ◽  
Substrate Cleavage ◽  
Caspase Substrate

scholarly journals Requirements for proteolysis during apoptosis.

1997 ◽  
Vol 17 (11) ◽  
pp. 6502-6507 ◽  
Author(s):  
D L Vaux ◽  
S Wilhelm ◽  
G Häcker
Keyword(s):  
Dna Cleavage ◽  
Morphological Changes ◽  
Cysteine Proteases ◽  
Dna Degradation ◽  
Effector Proteins ◽  
Proteinase K ◽  
Caspase Activation ◽  
Caspase Substrate ◽  
Do So

The key effector proteins of apoptosis are a family of cysteine proteases termed caspases. Following activation of caspases, biochemical events occur that lead to DNA degradation and the characteristic morphological changes associated with apoptosis. Here we show that cytoplasmic extracts activated in vitro by proteinase K were able to cleave the caspase substrate DEVD-7-amino-4-methylcoumarin, while neither proteinase K nor nonactivated extracts were able to do so alone. Caspase-like activity was inhibited by the specific caspase inhibitor DEVD-aldehyde and by the protease inhibitor iodoacetamide, but not by N-ethylmaleimide. When added to isolated nuclei, the activated extracts caused internucleosomal DNA degradation and morphological changes typical of apoptosis. As DNA cleavage and morphological changes could be inhibited by N-ethylmaleimide but not by iodoacetamide, we conclude that during apoptosis, caspase activation causes activation of another cytoplasmic enzyme that can be inhibited by N-ethylmaleimide. Activity of this enzyme is necessary for activation of endonucleases, DNA cleavage, and changes in nuclear morphology.

scholarly journals Unraveling oxidative stress response in the cestode parasite Echinococcus granulosus

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Martín Cancela ◽  
Jéssica A. Paes ◽  
Hercules Moura ◽  
John R. Barr ◽  
Arnaldo Zaha ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Echinococcus Granulosus ◽  
26S Proteasome ◽  
Cystic Hydatid Disease ◽  
Lamin B ◽  
Treatment And Prevention ◽  
Molecular Events ◽  
Antioxidant Proteins ◽  
Caspase Substrate ◽  
Cystic Hydatid

Abstract Cystic hydatid disease (CHD) is a worldwide neglected zoonotic disease caused by Echinococcus granulosus. The parasite is well adapted to its host by producing protective molecules that modulate host immune response. An unexplored issue associated with the parasite’s persistence in its host is how the organism can survive the oxidative stress resulting from parasite endogenous metabolism and host defenses. Here, we used hydrogen peroxide (H2O2) to induce oxidative stress in E. granulosus protoescoleces (PSCs) to identify molecular pathways and antioxidant responses during H2O2 exposure. Using proteomics, we identified 550 unique proteins; including 474 in H2O2-exposed PSCs (H-PSCs) samples and 515 in non-exposed PSCs (C-PSCs) samples. Larger amounts of antioxidant proteins, including GSTs and novel carbonyl detoxifying enzymes, such as aldo-keto reductase and carbonyl reductase, were detected after H2O2 exposure. Increased concentrations of caspase-3 and cathepsin-D proteases and components of the 26S proteasome were also detected in H-PSCs. Reduction of lamin-B and other caspase-substrate, such as filamin, in H-PSCs suggested that molecular events related to early apoptosis were also induced. We present data that describe proteins expressed in response to oxidative stress in a metazoan parasite, including novel antioxidant enzymes and targets with potential application to treatment and prevention of CHD.

Application of pharmacodynamic assays in a phase Ia trial of Apo2L/TRAIL in patients with advanced tumors

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 3535-3535 ◽  
Author(s):  
Y. Pan ◽  
R. Xu ◽  
M. Peach ◽  
C. Huang ◽  
D. Branstetter ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Complete Analysis ◽  
Serum Samples ◽  
Signaling Complex ◽  
Apoptotic Markers ◽  
Effector Caspase ◽  
Luminescent Signal ◽  
Caspase Substrate ◽  
Dose Dependent ◽  
Advanced Tumors

3535 Background: Recombinant human (rh) Apo2L/TRAIL is a dual pro-poptotic receptor agonist (PARA) that induces apoptosis by binding to pro-apoptotic receptors DR4 and DR5, which recruit a death inducing signaling complex upon ligand binding. This results in activation of the effector caspase 3/7, that subsequently cleaves intracellular substrates to execute cellular apoptosis. A Phase1a trial is underway to evaluate the safety and tolerability of rhApo2L/TRAIL in patients with advanced tumors. The aim of this study was to develop and validate high-throughput pharmacodynamic assays to monitor rhApo2L/TRAIL activity in easily accessible patient samples such as serum. Methods: To monitor rhApo2L/TRAIL activity in patients, we optimized assays to measure the release of the apoptotic markers caspase 3/7, cytokeratin 18 (CK18), and genomic DNA (gDNA) in serum. Serum caspase 3/7 levels were monitored using the Caspase Glo kit, which generates a luminescent signal upon cleavage of a caspase 3/7 substrate; cleavage of the caspase substrate CK18 was measured using an optimized form of the M30 ELISA assay; gDNA was measured using a β-actin-specific TaqMan real-time PCR assay. Mice bearing Colo205 xenografts were treated with rhApo2L/TRAIL and sera were collected and assayed for apoptotic markers. Upon validating these assays, we monitored the levels of apoptotic markers in cancer patients who received rhApo2L/TRAIL. Results: We detected transient increases in apoptotic markers in mouse sera 8–24 hr after treatment with rhApo2L/TRAIL. This increase was dose-dependent and correlated with active caspase 3 detected by IHC in Colo205 tumors. In the phase Ia study, increases in serum caspase 3/7 and gDNA levels were observed in >50% of colorectal, lung and sarcoma patients evaluated. Preliminary analyses show the percentage of increase correlates using both analytes and is dose-dependent. Conclusions: These findings support the use of serum-based pharmacodynamic assays as a means to monitor rhApo2L/TRAIL activity in patients with advanced tumors. A complete analysis of all patient serum samples from the ongoing phase Ia trial will be reported. No significant financial relationships to disclose.

Sign in / Sign up

Export Citation Format

Share Document