Information on EC 3.4.21.B12 - prostase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.21.B12
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
prostase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
proteolysis of polypeptides
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
226408-87-3
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9001-01-8
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
gene PRSS17
Q9JIS2
SwissProt
Manually annotated by BRENDA team
EMSP1
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(Abz)-SKGR-SLIGK(N-[2,4-dintrophenyl]ethylenediamine)-Asp-OH
?
show the reaction diagram
-
fluorescence quenched peptide spanning the PAR-2 activation site, is rapidly cleaved by KLK4
-
-
?
32 kDa enamelin + H2O
?
show the reaction diagram
-
five cleavage sites on the C-terminal sides of R180, Y205, H206, M219 and R262. Kallikrein 4 digestion of the 32 kDa enamelin generates nine major cleavage products. 12 h of digestion with KLK4, all of the 32 kDa enamelin has been cleaved. Some cleavage products persist after 48 h of digestion
-
-
?
52 kDa prostatic acid phosphatase + H2O
33 kDa prostatic acid phosphatase + 19 kDa fragment
show the reaction diagram
amelogenin + H2O
?
show the reaction diagram
-
-
-
-
?
amelogenin + H2O
amelogenin fragments
show the reaction diagram
benzoyl-L-Ile-L-Glu-Gly-L-Arg-4-nitroanilide + H2O
benzoyl-L-Ile-L-Glu-Gly-L-Arg + 4-nitroaniline
show the reaction diagram
benzyl-FVR-p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-FR-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-FR + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-LR-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-LR + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
casein + H2O
?
show the reaction diagram
zymograph assay
-
-
?
collagen I + H2O
?
show the reaction diagram
-
limited degradation
-
-
?
Collagen IV + H2O
?
show the reaction diagram
-
limited degradation
-
-
?
D-Pro-L-Phe-L-Arg-4-nitroanilide + H2O
D-Pro-L-Phe-L-Arg + 4-nitroaniline
show the reaction diagram
D-Val-L-Leu-L-Lys-4-nitroanilide + H2O
D-Val-L-Leu-L-Lys + 4-nitroaniline
show the reaction diagram
D-Val-Leu-Arg-4-nitroanilide + H2O
D-Val-Leu-Arg + 4-nitroaniline
show the reaction diagram
EKK-7-amido-4-methylcoumarin + H2O
EKK + 7-amino-4-methylcoumarin
show the reaction diagram
-
9% of the activity with VPR-7-amido-4-methylcoumarin
-
-
?
fibrinogen alpha-chain + H2O
?
show the reaction diagram
-
-
-
-
?
Gelatin + H2O
?
show the reaction diagram
GPR-7-amido-4-methylcoumarin + H2O
GPR + 7-amino-4-methylcoumarin
show the reaction diagram
-
20% of the activity with VPR-7-amido-4-methylcoumarin
-
-
?
human ephrin-B2 + H2O
?
show the reaction diagram
low activity with recombinant human enzyme and human substrate
-
-
?
murine ephrin-B2 + H2O
?
show the reaction diagram
the substrate harbors N-glycosylation sites its extracellular domain sequence, R27-R178, good activity with recombinant human enzyme and murine substrate, the primary enzyme cleavage site in murine ephrin-B2 is verified as located between extracellular domain residues Arg178 and N179
cleavage fragments, overview
-
?
plasma hepatocyte growth factor activator zymogen + H2O
mature HGF activator + ?
show the reaction diagram
polypeptide + H2O
peptides
show the reaction diagram
pro-prostate-specific antigen + H2O
activated prostate-specific antigen
show the reaction diagram
pro-urokinase-type plasminogen activator + H2O
activated urokinase-type plasminogen activator
show the reaction diagram
proform of KLK3 + H2O
?
show the reaction diagram
-
Ile-Leu-Ser-Arg-/-Ile-Val activation site
-
-
?
protease-activated receptor-1 + H2O
?
show the reaction diagram
-
KLK4 cleaves protease-activated receptor-1 at the activation site on intact cell surface
-
-
?
QAR-7-amido-4-methylcoumarin + H2O
QAR + 7-amino-4-methylcoumarin
show the reaction diagram
-
38% of the activity with VPR-7-amido-4-methylcoumarin
-
-
?
Tos-Gly-Pro-Arg-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin + H2O
tosyl-Gly-Pro-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
urokinase-type plasminogen activator + H2O
?
show the reaction diagram
-
cleaves after Pro-Arg-Phe-Lys
-
-
?
urokinase-type plasminogen activator receptor + H2
?
show the reaction diagram
-
cleavage occurs in D1-D2 linker sequence and, to a lesser extent, in D3 juxtamembrane domain
-
-
?
urokinase-type plasminogen activator receptor + H2O
?
show the reaction diagram
-
cleaves after Thr-Tyr-Ser-Arg as well as in the sequence Val-Gln-Tyr-Arg-/-Ser-Gly
-
-
?
VLK-7-amido-4-methylcoumarin + H2O
VLK + 7-amino-4-methylcoumarin
show the reaction diagram
-
60% of the activity with VPR-7-amido-4-methylcoumarin
-
-
?
VPR-7-amido-4-methylcoumarin + H2O
VPR + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
Z-FVR-p-nitroanilide + H2O
Z-FVR + p-nitroaniline
show the reaction diagram
-
-
-
-
?
Z-Phe-Val-Arg-p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
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
52 kDa prostatic acid phosphatase + H2O
33 kDa prostatic acid phosphatase + 19 kDa fragment
show the reaction diagram
-
enzyme may be involved in the physiological clearance of prostatic acid phosphatase
-
?
plasma hepatocyte growth factor activator zymogen + H2O
mature HGF activator + ?
show the reaction diagram
-
activation, hepatocyte growth factor is mainly secreted from fibroblasts as an inactive single-chain precursor, which lacks biological activity
-
-
?
polypeptide + H2O
peptides
show the reaction diagram
pro-prostate-specific antigen + H2O
activated prostate-specific antigen
show the reaction diagram
protease-activated receptor-1 + H2O
?
show the reaction diagram
-
KLK4 cleaves protease-activated receptor-1 at the activation site on intact cell surface
-
-
?
additional information
?
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Aprotinin
benzamidine
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93% inhibition at 10 mM
Co2+
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-
Nalpha-p-tosyl-L-lysine chloromethyl ketone
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85% inhibition at 1 mM; i.e. TLCK
Ni2+
-
-
siRNA
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mediates knockdown of endogenous KLK4 in LNCaP prostate cancer cells whereby inhibiting their proliferation
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Soybean trypsin inhibitor
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99% inhibition at 1 mg/ml
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Zn2+
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; enzyme activity is high at zinc concentrations below 0.001 mM and decreases to about 30% at 0.1 mM. Zinc affects the K4 active site via the salt-bridge formed between the N terminus and Asp194 required for a functional active site
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
androgens
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up-regulation of gene expression
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doxycycline
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progestins
recombinant pig enamelysin
can activate the recombinant zymogen in vitro
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Thermolysin
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additional information
no activation of recombinant zymogen by native KLK4 enzyme in vitro
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.008
(Abz)-SKGR-SLIGK(N-[2,4-dintrophenyl]ethylenediamine)-Asp-OH
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-
0.0469
benzyl-FVR-p-nitroanilide
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thermolysin activated recombinant KLK4
0.052
VLK-7-amido-4-methylcoumarin
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pH 8.0, 37C
0.049
VPR-7-amido-4-methylcoumarin
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pH 8.0, 37C
0.0207
Z-FVR-p-nitroanilide
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in 50 mM HEPES (pH 7.0), 150 mM NaCl, 10 mM CaCl2 buffer, at 25C
0.0207
Z-Phe-Val-Arg-p-nitroanilide
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monomeric K4
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
7.9
(Abz)-SKGR-SLIGK(N-[2,4-dintrophenyl]ethylenediamine)-Asp-OH
Homo sapiens
-
-
1.8
benzyl-FVR-p-nitroanilide
Homo sapiens
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thermolysin activated recombinant KLK4
0.0078
VLK-7-amido-4-methylcoumarin
Homo sapiens
-
pH 8.0, 37C
0.033
VPR-7-amido-4-methylcoumarin
Homo sapiens
-
pH 8.0, 37C
3.64
Z-FVR-p-nitroanilide
Homo sapiens
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in 50 mM HEPES (pH 7.0), 150 mM NaCl, 10 mM CaCl2 buffer, at 25C
3.64
Z-Phe-Val-Arg-p-nitroanilide
Homo sapiens
-
monomeric K4
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.1
Ni2+
Homo sapiens
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; in 50 mM HEPES (pH 7.0), 150 mM NaCl, 10 mM CaCl2 buffer, at 25C
0.016 - 0.0164
Zn2+
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.31
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purified recombinant enzyme, substrate H-D-Val-L-Leu-L-Lys-4-nitroanilide
0.67
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purified recombinant enzyme, substrate N-benzoyl-L-Ile-L-Glu-Gly-L-Arg-4-nitroanilide
1.12
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purified recombinant enzyme, substrate H-D-Pro-L-Phe-L-Arg-4-nitroanilide
1.42
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purified recombinant enzyme, substrate H-D-Val-Leu-Arg-4-nitroanilide
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6
assay at, substrate 20 kDa amelogenin
7.2
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 10.3
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7% of maximal activity at ph 5.0, 32% at pH 5.5, and 40% at pH 10.3
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
high expression level
Manually annotated by BRENDA team
low expression level
Manually annotated by BRENDA team
low expression level, not in fetal brain
Manually annotated by BRENDA team
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downregulated expression in efussions compared to solid tumors, higher expression as compared to malignant mesothelioma
Manually annotated by BRENDA team
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breast carcinoma cell line
Manually annotated by BRENDA team
low expression level
Manually annotated by BRENDA team
medium expression level
Manually annotated by BRENDA team
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KLK4 endometrial mRNA expression is greatest on days 0, 5, and 10 when compared with days 12, 15, and 17 of the estrous cycle and greater in cyclic compared with pregnant gilts
Manually annotated by BRENDA team
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co-expression of KLK4 and PAR-2
Manually annotated by BRENDA team
low expression level
Manually annotated by BRENDA team
medium expression level
Manually annotated by BRENDA team
-
mesothelioma cell of both peritoneal and pleural origin express K4, with downregulated expression in efussions compared to solid tumors
Manually annotated by BRENDA team
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of prostate cancer bone metastases
Manually annotated by BRENDA team
-
gene overexpression
Manually annotated by BRENDA team
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epithelial ovarian carcinoma cells
Manually annotated by BRENDA team
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co-expression of agonist and PAR-2 in primary prostate cancer (benign glands of benign prostatic hyperplasia, prostatic intraepithelial neoplasia and cancer) and prostate cancer bone metastasis
Manually annotated by BRENDA team
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KLK4 protein is significantly overexpressed in malignant prostate compared with normal prostate. No significant differences in expression levels among well or poorly differentiated tumors
Manually annotated by BRENDA team
low expression level
Manually annotated by BRENDA team
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KLK4 mRNA and protein is not significantly changed in SaOs2 cells treated with conditioned media from either PC-3 cells or LNCaP cells. KLK4 mRNA is clearly increased following direct co-culture of SaOs2 cells with either PC-3 orLNCaP cells
Manually annotated by BRENDA team
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seminal plasma, varying amounts of enzyme
Manually annotated by BRENDA team
medium expression level
Manually annotated by BRENDA team
high expression level
Manually annotated by BRENDA team
low expression level
Manually annotated by BRENDA team
high expression level
Manually annotated by BRENDA team
low expression level
Manually annotated by BRENDA team
-
varying amounts of enzyme
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
enzyme contains an amino-terminal pre-propeptide sequence, indicating a potential secretory function
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
21000
-
x * 21000, self-activated recombinant, chimeric enzyme, SDS-PAGE
23997
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1 * 25000, SDS-PAGE, 1 * 23997, MALDI-TOF mass spectrometry
24000
-
MALDI-TOF mass spectrometry
28000
x * 28000, deglycosylated enzyme, SDS-PAGE
30000
-
x * 30000, glycosylated form, x * 25000, deglycosylated form, SDS-PAGE
34000
-
active K4, Western blot analysis
40000
-
pro-K4, Western blot analysis
700000
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gel filtration, aggregate of four octamers
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
-
1 * 25000, SDS-PAGE, 1 * 23997, MALDI-TOF mass spectrometry
additional information
-
enzyme forms complexes with alpha1-antitrypsin, alpha2-antiplasmin and alpha2-macroglobulin
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by vapor diffusion in hanging drops. K4 crystallised in the presence of 10 mM Ni2+ reveals tetragonal space group P4. In the presence of Zn2+ it reveals tetragonal space group P41212. In the presence of 10 mM Co2+ it reveals monoclinic space group P21. The three different K4 crystal forms with bound Ni2+, Zn2, and Co2+ contain two, four and 16 crystallographically independent K4 molecules per asymmetric unit, respectively. K4 possesses an unusual 99-loop that creates a groove-like acidic S2 subsite; hanging drop vapour diffusion method using 20% PEG-MME 2000, 100 mM Tris (pH 8.5) and 10 mM NiCl2, or 100 mM HEPES (pH 6.5) and 100 mM NaCl with 1.6 M ammonium sulfate, or with 10 mM CoCl2, 100 mM Mes buffer (pH 6.0) and 1.6 M ammonium sulfate
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80C, phosphate buffer, pH 7.4
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
affinity purified
-
by Ni-NTA chromatography; nickel-nitrilotriacetic acid-Sepharose column chromatography
-
native enzyme from developing teeth; recombinant enzyme from 293 kidney cells
nickel-nitrilotriacetic acid-Sepharose column chromatography and benzamidine-Sepharose column chromatography
on Ni-NTA resin
-
recombinant chimeric enzyme after isolation from inclusion bodies and refolding
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
adenovirus-mediated ectopic expression of KLK4 in PC-3 cells and DU145 cells
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construction of a chimeric enzyme with exchange of the pro-piece of the zymogen for that of prostate-specific antigen PSA to create an activation site suceptible for trypsin-type proteases, expression in Escherichia coli strain BL21(DE3)
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DNA and amino acid sequence determination and analysis
DNA sequence determination and analysis, chromosome mapping and genomic organization
-
DNA sequence determination and analysis, chromosome mapping and genomic organization, promotor analysis
expressed in Escherichia coli M15(pREP4) cells and in insect cells (nur hk5)
expressed in Escherichia coli; into the BamH/HindIII restriction sites of the pQE30 vector and expressed in Escherichia coli M15[pREP4] cells
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expressed in Rattus norvegicus and Mus musculus
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expression in Escherichia coli strain AD 494 (DE3) with and without the pro-peptide; expression in Spodoptera frugiperda SF9 cells via transfection with baculovirus; expression of inactive zymogen in human 293 kidney cells
gene KLK4, coupling of the coding sequence of mature enzyme to the pro-domain of prostate specific antigen at the 5' end, and V5 and poly-histidine encoding sequences at the 3' end in plasmid pIBm recombinant expression as inactive zymogen in Spodoptera frugiperda insect cell line Sf9
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KLK4, 5, 6, and 7 full-length cDNA cloned into vector pRc/RSV. OV-MZ-6 ovarian cancer cells stably co-transfected with plasmids expressing K4-7. These OVKLK4+5+6+7 cells inoculated into the peritoneal cavity of nude mice
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KLK4-pIB/V5-His construct expressed in Sf9 cells. PC-3 cell stimulated with KLK4
-
overexpression of KLK4 in PC-3 cells
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
kallikrein expression is upregulated in prostate cancer cells
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E77A
-
the mutation completely abolishes the inhibitory effect of Zn2+ for hK4; zinc-independent
H25A
-
the mutation completely abolishes the inhibitory effect of Zn2+ for hK4; zinc-independent
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
isolation from inclusion bodies after recombinant expression in Escherichia coli, refolding with self-activation during the process
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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enzyme-specific sandwich-type immunoassay using a monoclonal antibody, detection limit is 0.02 microgram per liter, and less than 0.1% cross-reactivity with other human kallikreins
medicine