Previous studies have shown that K562 chronic myelogenous leukemia cells are resistant to induction of apoptosis by a variety of agents, including the topoisomerase II (topo II) poison etoposide, when examined 4 to 24 hours after treatment with an initiating stimulus. In the present study, the responses of K562 cells ... More
Activation and cleavage of caspase-3 in apoptosis induced by experimental cerebral ischemia.
AuthorsNamura S, Zhu J, Fink K, Endres M, Srinivasan A, Tomaselli KJ, Yuan J, Moskowitz MA
JournalJ Neurosci
PubMed ID9570797
'We examined the expression, activation, and cellular localization of caspase-3 (CPP32) using immunohistochemistry, immunoblots, and cleavage of the fluorogenic substrate N-benzyloxycarbonyl-Asp-Glu-Val-Asp-7-amino-4-trifluoromethyl coumarin (zDEVD-afc) in adult mouse brain after temporary (2 hr) middle cerebral artery occlusion produced by filament insertion into the carotid artery. Immunoreactive caspase-3p32 but not its cleavage product ... More
Reduced susceptibility of nonobese diabetic mice to TNF-alpha and D-galactosamine-mediated hepatocellular apoptosis and lethality.
'Nonobese diabetic (NOD/LtJ or NOD) mice are resistant to doses of LPS and D-galactosamine that uniformly produce lethality in C57BL/6J (B6) mice (p < 0.01). Liver caspase-3-like activity, serum transaminase levels (both p < 0.05), and the numbers of apoptotic liver nuclei were also reduced in NOD compared with B6 ... More
Fluorometric and colorimetric detection of caspase activity associated with apoptosis.
AuthorsGurtu V, Kain SR, Zhang G
JournalAnal Biochem
PubMed ID9300088
'The caspase (ICE/CED-3) family of proteases has been implicated to play a crucial role in apoptosis. However, the mechanisms by which caspase activity mediates apoptosis are not fully understood. Progress in this area has been limited due to the lack of a convenient and reliable system to quantify these protease ... More
Inhibition of caspase activity does not prevent the signaling phase of apoptosis in prostate cancer cells.
AuthorsDenmeade SR, Lin XS, Tombal B, Isaacs JT
JournalProstate
PubMed ID10344216
'BACKGROUND: Caspases are a family of cysteine proteases capable of characteristically cleaving after an aspartic acid residue. Various members of the caspase family (e.g., caspases 8 and 9) have been implicated as critical initiators in the signaling phase, while others (e.g., caspases 3, 6, and 7) have been implicated in ... More
E2F1 mediates death of B-amyloid-treated cortical neurons in a manner independent of p53 and dependent on Bax and caspase 3.
AuthorsGiovanni A, Keramaris E, Morris EJ, Hou ST, O'Hare M, Dyson N, Robertson GS, Slack RS, Park DS
JournalJ Biol Chem
PubMed ID10766769
'Although B-amyloid (AB) is suggested to play an important role in Alzheimer''s disease, the mechanisms that control AB-evoked toxicity are unclear. We demonstrated previously that the cell cycle-related cyclin-dependent kinase 4/6/retinoblastoma protein pathway is required for AB-mediated death. However, the downstream target(s) of this pathway are unknown. We show here ... More
The role of ARK in stress-induced apoptosis in Drosophila cells.
AuthorsZimmermann KC, Ricci JE, Droin NM, Green DR
JournalJ Cell Biol
PubMed ID11901172
'The molecular mechanisms of apoptosis are highly conserved throughout evolution. The homologs of genes essential for apoptosis in Caenorhabditis elegans and Drosophila melanogaster have been shown to be important for apoptosis in mammalian systems. Although a homologue for CED-4/apoptotic protease-activating factor (Apaf)-1 has been described in Drosophila, its exact function ... More
Possible involvement of cytochrome c release and sequential activation of caspases in ceramide-induced apoptosis in SK-N-MC cells.
AuthorsIto A, Uehara T, Tokumitsu A, Okuma Y, Nomura Y
JournalBiochim Biophys Acta
PubMed ID10590315
'Ceramide is characterized as a second messenger of apoptosis induced by various agents such as tumor necrosis factor (TNF-alpha), Fas ligand, hydrogen peroxide, heat shock and ionizing radiation. In this study, we investigated the mechanism of ceramide-induced apoptosis using a human neuroblastoma cell line, SK-N-MC. N-Acetyl-sphingosine (C2-ceramide), a cell-permeable ceramide ... More
Removal of the pro-domain does not affect the conformation of the procaspase-3 dimer.
AuthorsPop C, Chen YR, Smith B, Bose K, Bobay B, Tripathy A, Franzen S, Clark AC
JournalBiochemistry
PubMed ID11714276
'We have investigated the oligomeric properties of procaspase-3 and a mutant that lacks the pro-domain (called pro-less variant). In addition, we have examined the interactions of the 28 amino acid pro-peptide when added in trans to the pro-less variant. By sedimentation equilibrium studies, we have found that procapase-3 is a ... More
HSP27 inhibits cytochrome c-dependent activation of procaspase-9.
AuthorsGarrido C, Bruey JM, Fromentin A, Hammann A, Arrigo AP, Solary E
JournalFASEB J
PubMed ID10544189
'We have previously shown that the small heat shock protein HSP27 inhibited apoptotic pathways triggered by a variety of stimuli in mammalian cells. The present study demonstrates that HSP27 overexpression decreases U937 human leukemic cell sensitivity to etoposide-induced cytotoxicity by preventing apoptosis. As observed for Bcl-2, HSP27 overexpression delays poly(ADP-ribose)polymerase ... More
Fas-mediated apoptosis in mouse hepatocytes involves the processing and activation of caspases.
AuthorsJones RA, Johnson VL, Buck NR, Dobrota M, Hinton RH, Chow SC, Kass GE
JournalHepatology
PubMed ID9620337
'The mechanism of Fas antigen-induced hepatocyte apoptosis was investigated. Using a monoclonal antibody directed against the Fas antigen, apoptosis was induced in freshly isolated murine hepatocytes within 90 minutes of antibody addition as assessed by plasma membrane bleb formation, chromatin condensation, and DNA fragmentation. Pretreatment of the cells with the ... More
Involvement of caspase 3 in apoptotic death of cortical neurons evoked by DNA damage.
AuthorsKeramaris E, Stefanis L, MacLaurin J, Harada N, Takaku K, Ishikawa T, Taketo MM, Robertson GS, Nicholson DW, Slack RS, Park DS
JournalMol Cell Neurosci
PubMed ID10845773
'Previous reports have shown that DNA-damage-evoked death of embryonic cortical neurons is delayed by general caspase inhibitors and is accompanied by an increase in DEVD-AFC cleavage activity. We show here that this cleavage activity is lacking in camptothecin-treated caspase 3-deficient neurons. Moreover, we report that death of camptothecin-treated caspase 3-deficient ... More
Annexin-mediated Ca2+ influx regulates growth plate chondrocyte maturation and apoptosis.
AuthorsWang W, Xu J, Kirsch T
JournalJ Biol Chem
PubMed ID12446691
'Maturation of epiphyseal growth plate chondrocytes plays an important role in endochondral bone formation. Previously, we demonstrated that retinoic acid (RA) treatment stimulated annexin-mediated Ca(2+) influx into growth plate chondrocytes leading to a significant increase in cytosolic Ca(2+), whereas K-201, a specific annexin Ca(2+) channel blocker, inhibited this increase markedly. ... More
Defective cytochrome c-dependent caspase activation in ovarian cancer cell lines due to diminished or absent apoptotic protease activating factor-1 activity.
Authors Wolf B B; Schuler M; Li W; Eggers-Sedlet B; Lee W; Tailor P; Fitzgerald P; Mills G B; Green D R;
JournalJ Biol Chem
PubMed ID11429402
'Apoptosis via the mitochondrial pathway requires release of cytochrome c into the cytosol to initiate formation of an oligomeric apoptotic protease-activating factor-1 (APAF-1) apoptosome. The apoptosome recruits and activates caspase-9, which in turn activates caspase-3 and -7, which then kill the cell by proteolysis. Because inactivation of this pathway may ... More
Recombinant caspase-3 expressed in Pichia pastoris is fully activated and kinetically indistinguishable from the native enzyme.
AuthorsSun J, Bottomley SP, Kumar S, Bird PI
JournalBiochem Biophys Res Commun
PubMed ID9325193
'Intracellular cysteine proteinases (caspases) play key roles in inflammation and apoptosis. Recombinant caspases are typically produced in Escherichia coli expression systems with the attendant problems of solubilization, re-folding and activation of the protease. Here we describe the expression of hexahistidine-tagged caspase-3 (CPP32/Yama/Apopain) in the methylotropic yeast Pichia pastoris, and the ... More
Caspase-3 activity is present in cerebrospinal fluid from patients with traumatic brain injury.
AuthorsHärter L, Keel M, Hentze H, Leist M, Ertel W
JournalJ Neuroimmunol
PubMed ID11730942
Loss of neurons after traumatic brain injury (TBI) might involve dysregulated apoptosis. Activation of caspase-3 is one hallmark of apoptosis. Therefore, caspase-3 activity (cleavage of DEVD-afc) was measured in cerebrospinal fluid (CSF) samples (n=113) from 27 patients with TBI at day 1 to 14 after trauma. Caspase-3 activity was detected ... More
Structural basis for the inhibition of caspase-3 by XIAP.
AuthorsRiedl SJ, Renatus M, Schwarzenbacher R, Zhou Q, Sun C, Fesik SW, Liddington RC, Salvesen GS
JournalCell
PubMed ID11257232
The molecular mechanism(s) that regulate apoptosis by caspase inhibition remain poorly understood. The main endogenous inhibitors are members of the IAP family and are exemplified by XIAP, which regulates the initiator caspase-9, and the executioner caspases-3 and -7. We report the crystal structure of the second BIR domain of XIAP ... More
Disparate cleavage of poly-(ADP-ribose)-polymerase (PARP) and a synthetic tetrapeptide, DEVD, by apoptotic cells.
AuthorsYu K, Kennedy CA, O'Neill MM, Barton RW, Tatake RJ
JournalApoptosis
PubMed ID11388664
In the present investigations, we have shown differential cleavage of cellular PARP and a caspase 3-selective synthetic tetrapeptide substrate, Z-DEVD-AFC or Ac-DEVD-AMC using a T lymphoblastoid cell line Jurkat, and its variant clone E6.1(J-E6). Anti-Fas antibody-mediated apoptosis resulted in DNA fragmentation and PARP cleavage in both Jurkat and J-E6 cells. ... More
Mechanism of nitric oxide-induced apoptosis in human neuroblastoma SH-SY5Y cells.
AuthorsMoriya R, Uehara T, Nomura Y
JournalFEBS Lett
PubMed ID11078888
We have attempted to elucidate the precise mechanism of nitric oxide (NO)-induced apoptotic neuronal cell death. Enzymatic cleavages of DEVD-AFC, VDVAD-AFC, and LEHD-AFC (specific substrates for caspase-3-like protease (caspase-3 and -7), caspase-2, and caspase-9, respectively) were observed by treatment with NO. Western blot analysis showed that pro-forms of caspase-2, -3, ... More
BAX-induced cell death may not require interleukin 1 beta-converting enzyme-like proteases.
AuthorsXiang J, Chao DT, Korsmeyer SJ
JournalProc Natl Acad Sci U S A
PubMed ID8962091
Expression of BAX, without another death stimulus, proved sufficient to induce a common pathway of apoptosis. This included the activation of interleukin 1 beta-converting enzyme (ICE)-like proteases with cleavage of the endogenous substrates poly(ADP ribose) polymerase and D4-GDI (GDP dissociation inhibitor for the rho family), as well as the fluorogenic ... More
Prolonged therapeutic window for ischemic brain damage caused by delayed caspase activation.
AuthorsFink K, Zhu J, Namura S, Shimizu-Sasamata M, Endres M, Ma J, Dalkara T, Yuan J, Moskowitz MA
JournalJ Cereb Blood Flow Metab
PubMed ID9778183
Apoptotic cell death is prominent in neurodegenerative disorders, such as Alzheimer's disease and Huntington's disease, and is found in cerebral ischemia. Using a murine model of delayed cell death, we determined that cleavage of zDEVD-amino-4-trifluoromethyl coumarin (zDEVD-afc) in brain homogenate, a measure of caspase activation, increased initially 9 hours after ... More
Caspases: decoders of apoptotic signals during mammary involution. Caspase activation during involution.
AuthorsMarti A, Graber H, Lazar H, Ritter PM, Baltzer A, Srinivasan A, Jaggi R
JournalAdv Exp Med Biol
PubMed ID10959427
At weaning most of the alveolar epithelial cells in the mammary gland die by apoptosis and are removed by phagocytosis. Caspases are a family of aspartate specific cysteine proteases. Activation of caspases is generally thought to represent a major and irreversible event in the apoptotic process. We analyzed caspase expression ... More
Induction of apoptosis by a novel copper-based anticancer compound, casiopeina II, in L1210 murine leukaemia and CH1 human ovarian carcinoma cells.
AuthorsDe Vizcaya-Ruiz A, Rivero-Muller A, Ruiz-Ramirez L, Kass GE, Kelland LR, Orr RM, Dobrota M
JournalToxicol In Vitro
PubMed ID10699355
The activity of casiopeina II [Cu(1,4-dimethyl-1, 10-phenanthroline)(glycine)NO(3)], a novel anticancer agent, was tested in two cell lines, L1210 murine leukaemia, CH1 human ovarian carcinoma, cisplatin-resistant and sensitive. Exposure of the cells to a range of concentrations of casiopeina II indicates that this copper complex kills cells by apoptosis and necrosis. ... More
Pivotal role of oligomerization in expanded polyglutamine neurodegenerative disorders.
AuthorsSánchez I, Mahlke C, Yuan J
JournalNature
PubMed ID12540902
The expansion of a CAG repeat coding for polyglutamine in otherwise unrelated gene products is central to eight neurodegenerative disorders including Huntington's disease. It has been well documented that expanded polyglutamine fragments, cleaved from their respective full-length proteins, form microscopically visible aggregates in affected individuals and in transgenic mice. The ... More
Spontaneous in contrast to CD95-induced neutrophil apoptosis is independent of caspase activity.
AuthorsHärter L, Keel M, Hentze H, Steckholzer U, Ungethüm U, Trentz O, Ertel W
JournalJ Trauma
PubMed ID11426111
BACKGROUND: The influence of caspase inhibitors on spontaneous and on CD95-triggered apoptosis was investigated in neutrophils from healthy volunteers and compared with neutrophils from patients with severe sepsis. METHODS: To further elucidate the mechanisms of neutrophil apoptosis, isolated neutrophils from healthy volunteers (n = 9) were either stimulated with the ... More
Spontaneous human monocyte apoptosis utilizes a caspase-3-dependent pathway that is blocked by endotoxin and is independent of caspase-1.
AuthorsFahy RJ, Doseff AI, Wewers MD
JournalJ Immunol
PubMed ID10438906
Apoptosis is an important mechanism for regulating the numbers of monocytes and macrophages. Caspases (cysteine-aspartate-specific proteases) are key molecules in apoptosis and require proteolytic removal of prodomains for activity. Caspase-1 and caspase-3 have both been connected to apoptosis in other model systems. The present study attempted to delineate what role ... More
DEDD regulates degradation of intermediate filaments during apoptosis.
AuthorsLee JC, Schickling O, Stegh AH, Oshima RG, Dinsdale D, Cohen GM, Peter ME
JournalJ Cell Biol
PubMed ID12235123
Apoptosis depends critically on regulated cytoskeletal reorganization events in a cell. We demonstrate that death effector domain containing DNA binding protein (DEDD), a highly conserved and ubiquitous death effector domain containing protein, exists predominantly as mono- or diubiquitinated, and that diubiquitinated DEDD interacts with both the K8/18 intermediate filament network ... More
Activation of pro-death Bcl-2 family proteins and mitochondria apoptosis pathway in tumor necrosis factor-alpha-induced liver injury.
AuthorsZhao Y, Li S, Childs EE, Kuharsky DK, Yin XM
JournalJ Biol Chem
PubMed ID11369777
Tumor necrosis factor-alpha (TNFalpha)-induced cytotoxicity contributes to the pathogenesis in inflammatory and immune responses. Here, we studied the role of pro-death Bcl-2 family proteins and the mitochondria apoptosis pathway in the development of TNFalpha-induced hepatic injury during endotoxemia. After treating mice with lipopolysaccharide or TNFalpha in the presence of d-galactosamine, ... More
Apoptosis induced by dithiothreitol in HL-60 cells shows early activation of caspase 3 and is independent of mitochondria.
AuthorsTartier L, McCarey YL, Biaglow JE, Kochevar IE, Held KD
JournalCell Death Differ
PubMed ID11279547
Previous studies have shown that under certain conditions some thiol-containing compounds can cause apoptosis in a number of different cell lines. Herein, we investigated the apoptotic pathways in HL-60 cells triggered by dithiothreitol (DTT), used as a model thiol compound, and tested the hypothesis that thiols cause apoptosis via production ... More
Apoptosis induced by immunotoxins used in the treatment of hematologic malignancies.
AuthorsKeppler-Hafkemeyer A, Kreitman RJ, Pastan I
JournalInt J Cancer
PubMed ID10861457
The recombinant immunotoxins anti-Tac(Fv)-PE38 (LMB-2), targeting the interleukin-2 receptor alpha subunit (IL-2Ralpha, Tac or CD25), and RFB4(dsFv)-PE38 (BL22), targeting CD22, are being evaluated in clinical trials as treatment for hematologic malignancies. The toxin moiety Pseudomonas exotoxin A (PE) of these recombinant molecules leads to the arrest of protein synthesis due ... More
Activation of caspase-3 and apoptosis in cerebellar granule cells.
AuthorsMarks N, Berg MJ, Guidotti A, Saito M
JournalJ Neurosci Res
PubMed ID9590441
Caspase-3 activity increased dramatically in cytosolic extracts of rat cerebellar granule cells exposed to apoptotic conditions (basal medium Eagle (BME) containing 5 mM K+ without serum) when assayed with Ac-DEVD-amc, but not with Ac-YVAD-afc, a preferred substrate for caspase-1. This provided a basis to examine relationships between enzyme activity and ... More
Changes of caspase activities involved in apoptosis of a macrophage-like cell line J774.1/JA-4 treated with lipopolysaccharide (LPS) and cycloheximide.
AuthorsKarahashi H, Amano F
JournalBiol Pharm Bull
PubMed ID10706374
The addition of lipopolysaccharide (LPS) together with cycloheximide (CHX) induced apoptosis in a subline of a J774.1 macrophage-like cell line, JA-4, as judged by terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL)-staining and poly(adenosine 5'-diphosphate (ADP)-ribose) polymerase (PARP)-cleavage. Caspase activities were examined in these macrophages in vitro ... More
The mitochondrial permeability transition augments Fas-induced apoptosis in mouse hepatocytes.
AuthorsHatano E, Bradham CA, Stark A, Iimuro Y, Lemasters JJ, Brenner DA
JournalJ Biol Chem
PubMed ID10766806
Tumor necrosis factor-alpha receptor 1 and Fas recruit overlapping signaling pathways. To clarify the differences between tumor necrosis factor alpha (TNFalpha) and Fas pathways in hepatocyte apoptosis, primary mouse hepatocytes were treated with TNFalpha or an agonist anti-Fas antibody after infection with an adenovirus expressing an IkappaB superrepressor (Ad5IkappaB). Treatment ... More
Caspase-7 is activated during lovastatin-induced apoptosis of the prostate cancer cell line LNCaP.
AuthorsMarcelli M, Cunningham GR, Haidacher SJ, Padayatty SJ, Sturgis L, Kagan C, Denner L
JournalCancer Res
PubMed ID9426061
The goals of this work were to establish a reproducible and effective model of apoptosis in a cell line derived from advanced prostate cancer and to study the role of the caspase family of proteases in mediating apoptosis in this system. The study involved the use of the prostate cancer ... More
Role of caspases in immunotoxin-induced apoptosis of cancer cells.
AuthorsKeppler-Hafkemeyer A, Brinkmann U, Pastan I
JournalBiochemistry
PubMed ID9836586
Immunotoxins composed of antibodies linked to plant or bacterial toxins are being evaluated in the treatment of cancer. It is known that the toxin moieties of immunotoxins, including Pseudomonasexotoxin A (PE), diphtheria toxin, and ricin, are capable of inducing apoptosis. Since the efficiency of induction of apoptosis and the apoptosis ... More
Phosphorylated forms of activated caspases are present in cytosol from HL-60 cells during etoposide-induced apoptosis.
Treatment of HL-60 human leukemia cells with etoposide induces apoptotic cell death and activation of at least 18 electrophoretically distinct cysteine-dependent aspartate-directed protease (caspase) isoforms, several of which differ only in their isoelectric points. The purpose of the present study was to determine whether activated caspases are phosphorylated. Phosphatase treatment ... More
Presentation of nitric oxide regulates monocyte survival through effects on caspase-9 and caspase-3 activation.
AuthorsZeigler MM, Doseff AI, Galloway MF, Opalek JM, Nowicki PT, Zweier JL, Sen CK, Marsh CB
JournalJ Biol Chem
PubMed ID12566444
In the absence of survival factors, blood monocytes undergo spontaneous apoptosis, which involves the activation of caspase-3. Although nitric oxide can block caspase-3 activation and promote cell survival, it can also induce apoptosis. We hypothesized that nitrosothiols that promote protein S-nitrosylation would reduce caspase-3 activation and cell survival, whereas nitric ... More
Lack of correlation between caspase activation and caspase activity assays in paclitaxel-treated MCF-7 breast cancer cells.
AuthorsKottke TJ, Blajeski AL, Meng XW, Svingen PA, Ruchaud S, Mesner PW, Boerner SA, Samejima K, Henriquez NV, Chilcote TJ, Lord J, Salmon M, Earnshaw WC, Kaufmann SH
JournalJ Biol Chem
PubMed ID11677238
MCF-7 human breast cancer cells are widely utilized to study apoptotic processes. Recent studies demonstrated that these cells lack procaspase-3. In the present study, caspase activation and activity were examined in this cell line after treatment with the microtubule poison paclitaxel. When cells were harvested 72 h after the start ... More
Photodynamic therapy using Lu-Tex induces apoptosis in vitro, and its effect is potentiated by angiostatin in retinal capillary endothelial cells.
AuthorsRenno RZ, Delori FC, Holzer RA, Gragoudas ES, Miller JW
JournalInvest Ophthalmol Vis Sci
PubMed ID11053300
PURPOSE: To examine the effect of combining angiostatin with photodynamic therapy (PDT) using Lutetium Texaphyrin (Lu-Tex; Alcon, Fort Worth, TX) as a photosensitizer in bovine retinal capillary endothelial (BRCE) and retinal pigment epithelial (RPE) cells and to determine the mode of PDT-induced cell death in these cell lines. METHODS: Cultured ... More