Information on EC 3.4.22.B6 - cathepsin B-like protease

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.22.B6
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
cathepsin B-like protease
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
the enzyme does not readily hyrolyze benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin (which is cleaved by cathespsin B). The enzyme is not able to cleave synthetic substrates having an Arg in position P2
show the reaction diagram
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-
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
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-
-
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CAS REGISTRY NUMBER
COMMENTARY hide
37353-41-6
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
chili pepper
-
-
Manually annotated by BRENDA team
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G9TBU6
UniProt
Manually annotated by BRENDA team
female worms, three isozymes of HCB
-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
gene CPC
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
inhibition of the lysosomal cathepsin B-like protease blocks endocytosis causing cell death. Nicotinamide causes cytokinesis defects and cell cycle arrest at G2 phase
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-aminobenzoyl-Phe-Arg-Ala-(2,4-dinitrophenyl)-epsilon-NH2-lysine + H2O
?
show the reaction diagram
-
-
-
?
Arg-Arg-7-amido-4-methylcoumarin + H2O
Arg-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin + H2O
CBZ-Arg-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Arg-Arg-p-nitroanilide + H2O
benzyloxycarbonyl-Arg-Arg + p-nitroaniline
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
Boc-Val-Leu-Lys-7-amido-4-methylcoumarin + H2O
Boc-Val-Leu-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
?
hemoglobinase + H2O
?
show the reaction diagram
-
degradation
-
-
?
human IgG + H2O
?
show the reaction diagram
-
degradation
-
-
?
myristoylated-alanine-rich C-kinase + H2O
?
show the reaction diagram
-
specific degradation, the enzyme degrades the protein involved in actin cytoskeleton regulation
-
-
?
myristoylated-alanine-rich C-kinase substrate + H2O
?
show the reaction diagram
-
i.e. MARCKS, specific degradation
-
-
?
Phe-Arg-7-amido-4-methylcoumarin + H2O
Phe-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
protein rGSII-D88N + H2O
?
show the reaction diagram
-
legume lectin
-
-
?
Tyr-Val-Ala-Asp-7-amido-4-methylcoumarin + H2O
Tyr-Val-Ala-Asp + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
Z-Arg-Arg-7-amido-4-methylcoumarin + H2O
Z-Arg-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
Z-Gly-Pro-Arg-7-amido-4-methylcoumarin + H2O
Z-Gly-Pro-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
Z-Leu-Arg-7-amido-4-methylcoumarin + H2O
Z-Leu-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
G9TBU6
-
-
-
?
Z-Phe-Arg-7-amido-4-methylcoumarin + H2O
Z-Phe-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
myristoylated-alanine-rich C-kinase + H2O
?
show the reaction diagram
-
specific degradation, the enzyme degrades the protein involved in actin cytoskeleton regulation
-
-
?
additional information
?
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(1S,2S)-2-(((S)-1-((4-Guanidinobutyl)amino)-4-methyl-1-oxopentan-2-yl)carbamoyl)cyclopropanecarboxylic acid
-
i.e. E-64
-
acetyl-Leu-Val-lysinal
350 nM, 37°C, pH 4.5, 1 h, 15% inhibition
CA-074
CA074
CA074Me
-
specific inhibition
cathepsin L inhibitor 1
-
has a lower inhibitory effct on cathepsin B-like protease
Cbz-Phe-Gly-NHO-Bz-pMe
350 nM, 37°X, pH 4.5, 1 h, 87% inhibition
Cystatin
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K11002
-
morpholine urea-Phe-homoPhe-vinylsulphonylphenyl
L-transepoxysuccinyl-L-leucylamido-(4-guanidino)-butane
E64
Leishmania cysteine peptidase inhibitor 1
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N-(L-3-trans-propylcarbamoyl-oxirane-2-carbonyl)-L-Ile-L-Pro
-
CA074, irreversible inhibitor
N-(L-3-trans-propylcarbamoyloxirane-2-carbonyl)-Ile-Pro-OH
CA-074, irreversible inhibitor
N-(Na-carbonyl-Cpd-Gln-Ala-al)-Leu
elastatinal, irreversible inhibitor
N-trans-(epoxysuccinyl)-L-leucine 4-guanidinobutylamide
E-64, irreversible inhibitor
nicotinamide
complete inhibition at 25 mM
PS28
-
IC50: 0.01 mM
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PS44
-
IC50: 0.01 mM
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PS50
-
IC50: 0.005 mM
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Z-Phe-Ala-diazomethylketone
irreversible inhibitor
Z-Phe-Phe-diazomethylketone
irreversible inhibitor
Z-Phe-Tyr(tBu)-diazomethylketone
irreversible inhibitor
ZLIII115A
-
IC50: 0.01 mM
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ZLIII33A
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IC50: 0.0005
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ZLIII43A
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IC50: 0.002 mM
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additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
DTT
no activity in buffer lacking DTT
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.281 - 0.308
benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin
0.0102 - 0.0332
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
0.1733
N2-benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin
-
0.1234
Nalpha-benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.145
benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin
Leishmania major
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pH 5.5, 25°C, wild-type enzyme
1.05 - 7.6
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
3 - 6
CBZ-Arg-Arg-7-amido-4-methylcoumarin
Trypanosoma congolense
B2C323
-
1.09
CBZ-Phe-Arg-7-amido-4-methylcoumarin
Trypanosoma congolense
B2C323
-
3.49
N2-benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin
Trypanosoma congolense
B2C323
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1.09
Nalpha-benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
Trypanosoma congolense
B2C323
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000022 - 0.000035
CA074
0.000045 - 0.000186
K11002
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.01
PS28, PS44
Leishmania major
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IC50: 0.01 mM
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0.005
PS50
Leishmania major
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IC50: 0.005 mM
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0.01
ZLIII115A
Leishmania major
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IC50: 0.01 mM
-
0.0005
ZLIII33A
Leishmania major
-
IC50: 0.0005 mM
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0.002
ZLIII43A
Leishmania major
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IC50: 0.002 mM
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 6.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.89
-
sequence calculation, mature protein
5.42
-
sequence calculation, preproprotein
5.9
calculated for mature enzyme
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
high expression level
Manually annotated by BRENDA team
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expression in virulence
Manually annotated by BRENDA team
; expressed mainly in bloodstream form
Manually annotated by BRENDA team
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developing, HCB expression analysis during embryogenesis, overview
Manually annotated by BRENDA team
-
of femal adult worms
Manually annotated by BRENDA team
-
neurons
Manually annotated by BRENDA team
-
hippocampal
Manually annotated by BRENDA team
G9TBU6
-
Manually annotated by BRENDA team
-
in the sphere of yolk granules
Manually annotated by BRENDA team
-
three isozymes of HCB
Manually annotated by BRENDA team
G9TBU6
extracellular and intracellular sporozoites
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
the excretory/secretory enzyme is synthesized as preproprotein composed of a signal sequence, a proenzyme region , and a mature proteinase sequence
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
24000
-
x * 24000, 2-D electrophoresis
27400
-
x * 37800, about, preproprotein, sequence calculation, x * 27400, about, mature protein, sequence calculation
28500
x * 28500, calculated for mature enzyme, x * 32000, SDS-PAGE (glycosylated)
32000
x * 28500, calculated for mature enzyme, x * 32000, SDS-PAGE (glycosylated)
35000
x * 35000, x * 45000, gelatin SDS-PAGE digestion assay
37000
-
x * 37000, recombinant detagged enzyme, SDS-PAGE
37800
-
x * 37800, about, preproprotein, sequence calculation, x * 27400, about, mature protein, sequence calculation
42000
x * 42000, recombinant zymogen, SDS-PAGE
43000
x * 43000, SDS-PAGE, predicted: 36700 Da (with pro-region and without signal peptide), 29000 Da (without pro-region)
45000
x * 35000, x * 45000, gelatin SDS-PAGE digestion assay
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
the preproprotein is composed of a signal sequence of 17 residues, a proenzyme region of 76 amino acids, and a mature proteinase sequence of 251 amino acids in length
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
TbCatB/CA074 complex, hanging-drop method
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, immobilized metal ion affinity chromatography (Ni2+)
-
cation-exchange chromatography (Mono S)
cystatin affinity chromatography
from supernatant, immobilized metal ion affinity chromatography (Ni2+), ion-exchange chromatography (Mono-Q)
-
immobilized metal ion affinity chromatography (Ni2+)
recombinant GST-tagged enzyme from Escherichia coli strain BL21 to homogeneity, the tag is cleaved off by thrombin
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recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; expression in Pichia pastoris
cDNA library screening, DNA and amino acid sequence determination and analysis, phylogenetic analysis and tree, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3)
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construction of a intestinal cDNA library, DNA sequence determination and analysis of several genes of the cbl family, analysis of 686 cbl ESTs from USA or UK isolates containing 123 clusters, sequence and antigenic diversity, overview
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expressed in Pichia pastoris
expression of GST-tagged enzyme in Escherichia coli strain BL21
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gene AC-cathB-1, DNA and amino acid sequence determination and analysis, sequence comparisosns and phylogenetic analysis, quantitative real-time RT-PCR expression analysis; gene AC-cathB-2, DNA and amino acid sequence determination and analysis, sequence comparisosns and phylogenetic analysis, quantitative real-time RT-PCR expression analysis
His-tagged version expressed in Pichia pastoris
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His-tagged version expressed in Sf9 cell
recombinant expression in Pichia pastoris
recombinant expression of eimeripain fused to the fluorescent protein mCherry
G9TBU6
six-His-tagged protein is expressed in Pichia pastoris strain X-33; six-His-tagged protein is expressed in Pichia pastoris strain X-33
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
N97Q/N207Q
-
(putative) glycosylation sites
N97Q/N207Q/N100Q
-
(putative) glycosylation sites
N97Q/N207Q/N111Q
-
(putative) glycosylation sites
N97Q/N207Q/N111Q/N236Q
-
(putative) glycosylation sites
N97Q/N207Q/N236Q
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(putative) glycosylation sites
G234E
-
mutation increases activity towards benzyloxycarbonyl-Arg-ARg-7-amido-4-methylcoumarin, 8fold over the wild-type recombinant enzyme
N216D
-
predicted glycosylation site
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
TcoCBc1 elicits an important immune response in experimentally infected cattle. The protein is a potential therapeutic target and is a plausible antigen for Trypanosoma congolense diagnosis