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Information on EC 3.4.22.62 - caspase-9

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EC Tree
     3 Hydrolases
         3.4 Acting on peptide bonds (peptidases)
             3.4.22 Cysteine endopeptidases
                3.4.22.62 caspase-9
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This record set is specific for:
UNIPROT: P55211 not found.
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Word Map
The enzyme appears in viruses and cellular organisms
Reaction Schemes
strict requirement for an Asp residue at position P1 and with a marked preference for His at position P2. It has a preferred cleavage sequence of Leu-Gly-His-Asp-/-Xaa
Synonyms
caspase-9, caspase 9, casp9, casp-9, ice-lap6, casp9-gamma, apaf-3, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
casp9-gamma
APAF
-
-
-
-
Apaf-3
-
-
-
-
apoptotic protease activating factor 3
-
-
-
-
apoptotic protease Mch-6
-
-
-
-
C14.010
-
-
-
-
CASP-9
-
-
-
-
caspase 9
-
-
-
-
ICE-LAP6
-
-
-
-
ICE-like apoptotic protease 6
-
-
-
-
Mch6
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
180189-96-2
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetyl-Asp-Glu-Val-Asp-p-nitroanilide + H2O
acetyl-Asp-Glu-Val-Asp + p-nitroaniline
show the reaction diagram
i.e. Ac-DEVD-pNA, caspase-9/-3 activation in differentiated cells can be prevented by protein kinase C (PKC) and the mitogen activated protein kinase (MEK) signaling pathways
-
-
?
acetyl-DEVD-7-amido-4-trifluoromethylcoumarin + H2O
acetyl-DEVD + 7-amino-4-trifluoromethylcoumarin
show the reaction diagram
apoptosome inhibitors analyzed, caspase-9 activity indirectly measured by caspase-3 assay
-
-
?
acetyl-LEHD-7-amido-4-trifluoromethyl coumarin + H2O
acetyl-LEHD + 7-amino-4-trifluoromethyl coumarin
show the reaction diagram
acetyl-LEHD-7-amido-4-trifluoromethylcoumarin + H2O
acetyl-LEHD + 7-amino-4-trifluoromethylcoumarin
show the reaction diagram
acetyl-Leu-Glu-His-Asp-p-nitroanilide + H2O
acetyl-Leu-Glu-His-Asp + p-nitroaniline
show the reaction diagram
poly(ADP-ribose) polymerase + H2O
?
show the reaction diagram
-
-
-
?
pro-caspase-3 + H2O
caspase-3 + ?
show the reaction diagram
procaspase-3 + H2O
caspase-3 + ?
show the reaction diagram
-
-
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
pro-caspase-3 + H2O
caspase-3 + ?
show the reaction diagram
procaspase-3 + H2O
caspase-3 + ?
show the reaction diagram
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cd2+
anti-apoptotic cell survival function of cadmium, cadmium inhibits apoptosis induced by benzo[a]pyrene-7,8-diol-9,10-epoxide (BPDE) at non-cytotoxic concentrations, cadmium chloride pre-treatment of cells significantly inhibits caspase-9 activation in a dose dependent manner, 40% and 52% inhibition at 10 and 20 microM cadmium chloride, respectively
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
benzyloxycarbonyl-LEHD-fluoromethylketone
benzyloxycarbonyl-Leu-Glu-His-Asp-fluoromethylketone
i.e. z-LEHD-FMK, inhibitor concentration 100 microM
cadmium
anti-apoptotic cell survival function of cadmium, cadmium inhibits apoptosis induced by benzo[a]pyrene-7,8-diol-9,10-epoxide (BPDE) at non-cytotoxic concentrations, 40% and 52% inhibition at 10 and 20 microM cadmium chloride, respectively
N-(2-amino-2-oxoethyl)-1-[2-(2,4-dichlorophenyl)ethyl]-4-(3,3-diphenylpropyl)-N-[2-(2-fluorophenyl)ethyl]-3,7-dioxo-1,4-diazepane-5-carboxamide
5 microM for cell treatment, cells harvested after 24, 48, and 72 h, 51% inhibition of caspase-3 activity in SAOS-2 cells
N-(2-amino-2-oxoethyl)-1-[2-(2,4-dichlorophenyl)ethyl]-4-(3,3-diphenylpropyl)-N-[2-(2-methoxyphenyl)ethyl]-3,7-dioxo-1,4-diazepane-5-carboxamide
5 microM for cell treatment, cells harvested after 24, 48, and 72 h, 1% inhibition of caspase-3 activity in SAOS-2 cells
N-(2-amino-2-oxoethyl)-1-[2-(2,4-dichlorophenyl)ethyl]-4-(3,3-diphenylpropyl)-N-[2-(4-fluorophenyl)ethyl]-3,7-dioxo-1,4-diazepane-5-carboxamide
5 microM for cell treatment, cells harvested after 24, 48, and 72, 34% inhibition of caspase-3 activity in SAOS-2 cells
N-(2-amino-2-oxoethyl)-N-(2-cyclopropylethyl)-1-[2-(2,4-dichlorophenyl)ethyl]-4-(3,3-diphenylpropyl)-3,7-dioxo-1,4-diazepane-5-carboxamide
5 microM for cell treatment, cells harvested after 24, 48, and 72 h, 16% inhibition of caspase-3 activity in SAOS-2 cells
N-(2-amino-2-oxoethyl)-N-butyl-1-[2-(2,4-dichlorophenyl)ethyl]-4-(3,3-diphenylpropyl)-3,7-dioxo-1,4-diazepane-5-carboxamide
5 microM for cell treatment, cells harvested after 24, 48, and 72 h for caspase activity assay, 45% inhibition of caspase-3 activity in SAOS-2 cells
N-(2-amino-2-oxoethyl)-N-[2-(4-chlorophenyl)ethyl]-1-[2-(2,4-dichlorophenyl)ethyl]-4-(3,3-diphenylpropyl)-3,7-dioxo-1,4-diazepane-5-carboxamide
5 microM for cell treatment, cells harvested after 24, 48, and 72 h, 2% inhibition of caspase-3 activity in SAOS-2 cells
N-[2-(acetylamino)ethyl]-N-(2-amino-2-oxoethyl)-1-[2-(2,4-dichlorophenyl)ethyl]-4-(3,3-diphenylpropyl)-3,7-dioxo-1,4-diazepane-5-carboxamide
5 microM for cell treatment, cells harvested after 24, 48, and 72 h, 22% inhibition of caspase-3 activity in SAOS-2 cells
PGA-1
PGA-peptoid conjugate, inhibitor of apoptotic protease activating factor 1 (Apaf-1) coupled to poly-L-glutamic acid (PGA), at concentrations of 50 and 100 microM inhibition of caspase-3 activity, reaches values up to 100% at 50 microM drug after 48 h in HeLa-cells and and after 72 h in SAOS-2 cells
RQIKIWFQNRRMKWKKGG-N-[2-(2,4-dichlorophenyl)ethyl]glycyl-N-(3,3-diphenylpropyl)glycyl-N2-[2-(2,4-dichlorophenyl)ethyl]glycinamide
i.e. PEN-1, modified compound with peptide bridge, 5 microM for cell treatment, shows low inhibitory activity of caspase-3
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
adenosine
extracellular adenosine induces apoptosis by activating caspase-9 and caspase-3 in association with mitochondrial damage via A2a adenosine receptors, induction dose-dependent (1-20 mM) and time-dependent (24-72 h)
APAF-1
interaction with APAF-1 promotes auto-cleavage and activation of caspase-9
-
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.466 - 0.686
acetyl-LEHD-7-amido-4-trifluoromethyl coumarin
0.466 - 0.686
acetyl-LEHD-7-amido-4-trifluoromethylcoumarin
0.139
procaspase-3
caspase-9 holoenzyme
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
caspase activity assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
human fibroblast GM03349
Manually annotated by BRENDA team
cell line HF1A3, wild-type, dominant negative caspase-9 overexpressing
Manually annotated by BRENDA team
cell line HF1A3, wild-type, dominant negative caspase-9 overexpressing
Manually annotated by BRENDA team
gingivial cancer cells (Ca9-22 cells) containing a mutated p53 (mp53) protein by point mutation at codon 248
Manually annotated by BRENDA team
cells treated with 10 mg/ml recombinant Vibrio vulnificus cytolysin (rVVC) at 37°C for 4 h, cell viability assay
Manually annotated by BRENDA team
intact protein p53
Manually annotated by BRENDA team
human H460 NSCLC cells with and without cytochrome c/(d)ATP-induced activation of apoptosis
Manually annotated by BRENDA team
wild-type and SOD-antisense
Manually annotated by BRENDA team
caspase-9 genotyping
Manually annotated by BRENDA team
treated with doxycycline (2 microg/ml in PBS)
Manually annotated by BRENDA team
cells treated with 10 mg/ml recombinant Vibrio vulnificus cytolysin (rVVC) at 37°C for 4 h, cell viability assay
Manually annotated by BRENDA team
cells treated with 10 mg/ml recombinant Vibrio vulnificus cytolysin (rVVC) at 37°C for 4 h, cell viability assay
Manually annotated by BRENDA team
treated with different concentrations of marine sponge extracts of Polymastia janeirensis
Manually annotated by BRENDA team
treated with doxorubicin (2, 2.5, and 0.25 microg/ml in PBS), doxorubicin-induced cell death
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
phosphorylation of caspase-9 by DYRK1A involves co-localization to the nucleus
Manually annotated by BRENDA team
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
CASP9_HUMAN
416
0
46281
Swiss-Prot
other Location (Reliability: 3)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30000
x * 30000, caspase 9S, SDS-PAGE
35000
processed caspase-9 protein, SDS-PAGE
37000
processed caspase-9 protein, SDS-PAGE
46200
x * 46200, calculation from nucleotide sequence
47000
un-processed caspase-9 protein, SDS-PAGE
55000
full-length caspase-9, Western blot analysis
68000
observed molecular weight of caspase-9 by tandem mass spectrometry in cell lysates with and without cytochrome c/dATP treatment
69200
calculated molecular weight of caspase-9 by tandem mass spectrometry in cell lysates with and without cytochrome c/dATP treatment
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C287A
immunofluorescence staining of cells transiently transfected with vectors encoding caspase-9 point mutation
C403S
point mutation at C403 of caspase-9 impairs its activation mediated by H2O2, interaction with APAF-1 diminished through the abolition of disulfide formation, weaker association between cytochrome c and the C403S mutant than that between cytochrome c and wild-type caspase-9
T125A/C287A
site-directed mutagenesis, immunofluorescence staining of cells transiently transfected with vectors encoding caspase-9 double mutant
additional information
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
gel filtration
gel filtration, SDS-PAGE
recombinant His6-tagged enzyme
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
caspase-9 inactivated by mutation at the catalytic domain, generation of dominant negative caspase-9 overexpressing cell lines, expression in Escherichia coli, follicular lymphoma cells transformed with the lentiviral vector pWPI-IRES-GFP
caspase-9 splice variant Casp9-gamma contains only a caspase recruitment domain and lacks the catalytic domain, expression in 293T cells.Casp9-gamma does not promote apoptosis when overexpressed in 293T cells
expressed in Escherichia coli, recombinant protein, pcDNA3, pET28a, pEGFP vectors
naturally occuring variant caspase-9S, that is missing most of the large subunit of caspase-9
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
pharmacology
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Duan H.; Orth K.; Chinnaiyan A.M.; Poirier G.G.; Froelich C.J.; He W.W.; Dixit V.M.
ICE-LAP6, a novel member of the ICE/Ced-3 gene family, is activated by the cytotoxic T cell protease granzyme B
J. Biol. Chem.
271
16720-16724
1996
Homo sapiens (P55211)
Manually annotated by BRENDA team
Srinivasula, S.M.; Fernandes-Alnemri, T.; Zangrilli, J.; Robertson, N.; Armstrong, R.C.; Wang, L.; Trapani, J.A.; Tomaselli, K.J.; Litwack, G.; Alnemri E.S.
The Ced-3/interleukin 1beta converting enzyme-like homolog Mch6 and the lamin-cleaving enzyme Mch2alpha are substrates for the apoptotic mediator CPP32
J. Biol. Chem.
271
27099-27106
1996
Homo sapiens (P55211)
Manually annotated by BRENDA team
Seol, D.W.; Billiar, T.R.
A caspase-9 variant missing the catalytic site is an endogenous inhibitor of apoptosis
J. Biol. Chem.
274
2072-2076
1999
Homo sapiens (P55211), Homo sapiens
Manually annotated by BRENDA team
Wang, P.; Shi, T.; Ma, D.
Cloning of a novel human caspase-9 splice variant containing only the CARD domain
Life Sci.
79
934-940
2006
Homo sapiens (P55211), Homo sapiens
Manually annotated by BRENDA team
Yin, Q.; Park, H.H.; Chung, J.Y.; Lin, S.C.; Lo, Y.C.; da Graca, L.S.; Jiang, X.; Wu, H.
Caspase-9 holoenzyme is a specific and optimal procaspase-3 processing machine
Mol. Cell
22
259-268
2006
Homo sapiens (P55211)
Manually annotated by BRENDA team
Eeva, J.; Nuutinen, U.; Ropponen, A.; Maettoe, M.; Eray, M.; Pellinen, R.; Wahlfors, J.; Pelkonen, J.
The involvement of mitochondria and the caspase-9 activation pathway in rituximab-induced apoptosis in FL cells
Apoptosis
14
687-698
2009
Homo sapiens (P55211)
Manually annotated by BRENDA team
Day, T.W.; Huang, S.; Safa, A.R.
c-FLIP knockdown induces ligand-independent DR5-, FADD-, caspase-8-, and caspase-9-dependent apoptosis in breast cancer cells
Biochem. Pharmacol.
76
1694-1704
2008
Homo sapiens (P55211)
Manually annotated by BRENDA team
Yamakawa, N.; Takahashi, A.; Mori, E.; Imai, Y.; Furusawa, Y.; Ohnishi, K.; Kirita, T.; Ohnishi, T.
High LET radiation enhances apoptosis in mutated p53 cancer cells through Caspase-9 activation
Cancer Sci.
99
1455-1460
2008
Homo sapiens (P55211), Homo sapiens
Manually annotated by BRENDA team
Zuo, Y.; Xiang, B.; Yang, J.; Sun, X.; Wang, Y.; Cang, H.; Yi, J.
Oxidative modification of caspase-9 facilitates its activation via disulfide-mediated interaction with APAF-1
Cell Res.
19
449-457
2009
Homo sapiens (P55211)
Manually annotated by BRENDA team
Seifert, A.; Allan, L.A.; Clarke, P.R.
DYRK1A phosphorylates caspase 9 at an inhibitory site and is potently inhibited in human cells by harmine
FEBS J.
275
6268-6280
2008
Homo sapiens (P55211), Homo sapiens
Manually annotated by BRENDA team
Yiang, G.T.; Yu, Y.L.; Hu, S.C.; Chen, M.H.; Wang, J.J.; Wei, C.W.
PKC and MEK pathways inhibit caspase-9/-3-mediated cytotoxicity in differentiated cells
FEBS Lett.
582
881-885
2008
Homo sapiens (P55211)
Manually annotated by BRENDA team
Tichy, A.; Zaskodova, D.; Pejchal, J.; Rezacova, M.; Osterreicher, J.; Vavrova, J.; Cerman, J.
Gamma irradiation of human leukaemic cells HL-60 and MOLT-4 induces decrease in Mcl-1 and Bid, release of cytochrome c, and activation of caspase-8 and caspase-9
Int. J. Radiat. Biol.
84
523-530
2008
Homo sapiens (P55211), Homo sapiens
Manually annotated by BRENDA team
Conte da Frota, M.L.; Braganhol, E.; Delgado Canedo, A.; Klamt, F.; Apel, M.A.; Mothes, B.; Lerner, C.; Oliveira Battastini, A.M.; Henriques, A.T.; Fonseca Moreira, J.C.
Extracts of marine sponge Polymastia janeirensis induce oxidative cell death through a caspase-9 apoptotic pathway in human U138MG glioma cell line
Invest. New Drugs
27
440-446
2008
Homo sapiens (P55211), Homo sapiens
Manually annotated by BRENDA team
Yasuda, Y.; Saito, M.; Yamamura, T.; Yaguchi, T.; Nishizaki, T.
Extracellular adenosine induces apoptosis in Caco-2 human colonic cancer cells by activating caspase-9/-3 via A(2a) adenosine receptors
J. Gastroenterol.
44
56-65
2009
Homo sapiens (P55211), Homo sapiens
Manually annotated by BRENDA team
Mondragon, L.; Orzaez, M.; Sanclimens, G.; Moure, A.; Arminan, A.; Sepulveda, P.; Messeguer, A.; Vicent, M.J.; Perez-Paya, E.
Modulation of cellular apoptosis with apoptotic protease-activating factor 1 (Apaf-1) inhibitors
J. Med. Chem.
51
521-529
2008
Rattus norvegicus, Homo sapiens (P55211)
Manually annotated by BRENDA team
Andreoli, V.; Trecroci, F.; La Russa, A.; Valentino, P.; Condino, F.; Latorre, V.; Nistico, R.; Pirritano, D.; Del Giudice, F.; Canino, M.; Cittadella, R.; Quattrone, A.
CASP-9: A susceptibility locus for multiple sclerosis in Italy
J. Neuroimmunol.
210
100-103
2009
Homo sapiens (P55211), Homo sapiens
Manually annotated by BRENDA team
Checinska, A.; Giaccone, G.; Rodriguez, J.A.; Kruyt, F.A.; Jimenez, C.R.
Comparative proteomics analysis of caspase-9-protein complexes in untreated and cytochrome c/dATP stimulated lysates of NSCLC cells
J. Proteomics
72
575-585
2009
Homo sapiens (P55211)
Manually annotated by BRENDA team
Zhao, J.F.; Sun, A.H.; Ruan, P.; Zhao, X.H.; Lu, M.Q.; Yan, J.
Vibrio vulnificus cytolysin induces apoptosis in HUVEC, SGC-7901 and SMMC-7721 cells via caspase-9/3-dependent pathway
Microb. Pathog.
46
194-200
2009
Homo sapiens (P55211), Homo sapiens
Manually annotated by BRENDA team
Mukherjee, J.J.; Gupta, S.K.; Kumar, S.
Inhibition of benzopyrene-diol-epoxide (BPDE)-induced bax and caspase-9 by cadmium: role of mitogen activated protein kinase
Mutat. Res.
661
41-46
2009
Homo sapiens (P55211), Homo sapiens
Manually annotated by BRENDA team
Wei, X.; Li, Q.; Han, Z.; Lin, D.; Yu, P.
Differences in caspase-8 and -9 activity and sperm motility in infertile males of Li nationality in China
Int. J. Clin. Exp. Med.
8
4721-4726
2015
Homo sapiens (P55211), Homo sapiens
Manually annotated by BRENDA team
Diaconu, I.; Ballard, B.; Zhang, M.; Chen, Y.; West, J.; Dotti, G.; Savoldo, B.
Inducible caspase-9 selectively modulates the toxicities of CD19-specific chimeric antigen receptor-modified T cells
Mol. Ther.
25
580-592
2017
Homo sapiens (P55211)
Manually annotated by BRENDA team