Any feedback?
Please rate this page
(enzyme.php)
(0/150)

BRENDA support

BRENDA Home
show all | hide all No of entries

Information on EC 3.4.21.9 - enteropeptidase and Organism(s) Homo sapiens and UniProt Accession P98073

for references in articles please use BRENDA:EC3.4.21.9
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
     3 Hydrolases
         3.4 Acting on peptide bonds (peptidases)
             3.4.21 Serine endopeptidases
                3.4.21.9 enteropeptidase
Specify your search results
Select one or more organisms in this record: ?
This record set is specific for:
Homo sapiens
UNIPROT: P98073 not found.
Show additional data
Do not include text mining results
Include (text mining) results
Include results (AMENDA + additional results, but less precise)
Word Map
The taxonomic range for the selected organisms is: Homo sapiens
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
Activation of trypsinogen by selective cleavage of Lys6-/-Ile bond
Synonyms
enterokinase, enteropeptidase, human enteropeptidase, l-hep, enteropeptidase light chain, bovine enteropeptidase, enterokinase light chain, porcine enterokinase, bovine enterokinase light chain, tmprss15, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
EPL
catalytic subunit
L-HEK
catalytic subunit
enterokinase
enteropeptidase
enteropeptidase light chain
-
L-HEP
EP 118-1035
-
-
human enteropeptidase
-
-
L-HEP
natural enteropeptidase
-
-
peptidase, entero-
-
-
-
-
PRSS7
-
-
TMPRSS15
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
9014-74-8
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
fusion protein Trx/hEGF + H2O
?
show the reaction diagram
the fusion protein Trx/hEGF contains an inter-domain enteropeptidase recognition site
-
?
fusion protein Trx/hIL-13 + H2O
?
show the reaction diagram
the fusion protein Trx/hIL-13 contains an inter-domain enteropeptidase recognition site
-
?
GD4R-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
?
GDDDDK-4-nitroanilide + H2O
GDDDDK + 4-nitroaniline
show the reaction diagram
-
-
-
?
Gly-(L-Asp)4-L-Lys-2-naphthylamide + H2O
Gly-(L-Asp)4-L-Lys + 2-naphthylamine
show the reaction diagram
-
-
-
?
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide + H2O
Gly-Asp-Asp-Asp-Asp-Lys + 2-naphthylamine
show the reaction diagram
Gly-L-Asp-L-Asp-L-Asp-L-Asp-L-Lys-2-naphthylamide + H2O
Gly-L-Asp-L-Asp-L-Asp-L-Asp-L-Lys + 2-naphthylamine
show the reaction diagram
-
-
-
?
Melittin + H2O
?
show the reaction diagram
-
-
-
?
thiobenzyl benzyloxy-carbonyl-L-lysinate + H2O
?
show the reaction diagram
-
-
?
trypsinogen + H2O
trypsin
show the reaction diagram
trypsinogen + H2O
trypsin + ?
show the reaction diagram
Z-Lys-SBzl + H2O
?
show the reaction diagram
-
-
-
?
A4-mutant trypsinogen + H2O
?
show the reaction diagram
-
-
-
-
?
AAVERW-streptavidin-R-phycoerythrin conjugate + H2O
AAVERW + streptavidin-R-phycoerythrin conjugate
show the reaction diagram
-
peptide conjugated to fluorescent probe streptavidin-conjugated phycoerythrin
-
-
?
APFDDDDKIVGG + H2O
?
show the reaction diagram
-
N-terminal dodecapeptides of human cationic trypsinogen
-
-
?
benzoyl-Arg ethyl ester + H2O
benzoyl-Arg + ethanol
show the reaction diagram
-
-
-
-
?
benzyl-L-Arg-2-naphthylamide + H2O
?
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-L-alanine-X-L-arginine-p-nitroanilide + H2O
p-nitroaniline + benzyloxycarbonyl-L-alanine-X-L-arginine
show the reaction diagram
-
Z-Ala-X-Arg-pNA
-
-
?
DDDK-SA-PE + H2O
DDDK + SA-PE
show the reaction diagram
-
Asp-Asp-Asp-Asp-Lys-fluorescent probe streptavidin-conjugated phycoerythrin
-
-
?
DDRRAG-SA-PE + H2O
DDRRAG + SA-PE
show the reaction diagram
-
peptide conjugated to fluorescent probe streptavidin-conjugated phycoerythrin
-
-
?
DRARVW-SA-PE + H2O
DRARVW + SA-PE
show the reaction diagram
-
peptide conjugated to fluorescent probe streptavidin-conjugated phycoerythrin
-
-
?
EYDRQL-SA-PE + H2O
EYDRQL + SA-PE
show the reaction diagram
-
peptide conjugated to fluorescent probe streptavidin-conjugated phycoerythrin
-
-
?
formyl-Ala-Phe-Arg-4-nitroanilide + H2O
4-nitroaniline + formyl-Ala-Phe-Arg
show the reaction diagram
-
-
-
-
?
formyl-Ala-Phe-Arg-4-nitroanilide + H2O
?
show the reaction diagram
-
a chromogenic peptide substrates
-
-
?
formyl-Ala-Phe-Lys-4-nitroanilide + H2O
4-nitroaniline + formyl-Ala-Phe-Lys
show the reaction diagram
-
-
-
-
?
formyl-Ala-Phe-Lys-4-nitroanilide + H2O
?
show the reaction diagram
-
a chromogenic peptide substrates
-
-
?
G5DKF(NO2)G + H2O
?
show the reaction diagram
-
-
-
-
?
GD4K 2-naphthylamide + H2O
?
show the reaction diagram
-
-
-
-
?
GD4K 7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
GD4KF(NO2)G + H2O
?
show the reaction diagram
-
-
-
-
?
GDDDDK-2-naphthylamide + H2O
GDDDDK + 2-naphthylamine
show the reaction diagram
-
-
-
-
?
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide + H2O
Gly-Asp-Asp-Asp-Asp-Lys + 2-naphthylamine
show the reaction diagram
glycyl-tetra-L-aspartyl-L-lysine beta-naphthylamide + H2O
glycyl-tetra-L-aspartyl-L-lysine + beta-naphthylamine
show the reaction diagram
-
-
-
-
?
human cationic trypsinogen + H2O
?
show the reaction diagram
-
-
-
-
?
LTAEEKA + H2O
?
show the reaction diagram
-
-
-
-
?
LTAEEKAAV + H2O
?
show the reaction diagram
-
-
-
-
?
MHGERM-SA-PE + H2O
MHGERM + SA-PE
show the reaction diagram
-
peptide conjugated to fluorescent probe streptavidin-conjugated phycoerythrin
-
-
?
MSGERM-SA-PE + H2O
MSGERM + SA-PE
show the reaction diagram
-
peptide conjugated to fluorescent probe streptavidin-conjugated phycoerythrin
-
-
?
N-alpha-benzyloxycarbonyl-L-lysine thiobenzyl ester + H2O
N-alpha-benzyloxycarbonyl-L-lysine + benzylmercaptane
show the reaction diagram
-
-
-
-
?
N-alpha-benzyloxycarbonyl-L-lysine thiobenzyl ester + H2O
N-alpha-benzyloxycarbonyl-L-lysine + phenylmethanethiol
show the reaction diagram
-
-
-
-
?
Nalpha-acetyltrypsinogen + H2O
Nalpha-acetylactivation peptides + trypsin
show the reaction diagram
-
-
-
?
protein C inhibitor + H2O
?
show the reaction diagram
-
-
-
-
?
SERAAAG-SA-PE + H2O
SERAAAG + SA-PE
show the reaction diagram
-
peptide conjugated to fluorescent probe streptavidin-conjugated phycoerythrin
-
-
?
SGDRMW-SA-PE + H2O
SGDRMW + SA-PE
show the reaction diagram
-
peptide conjugated to fluorescent probe streptavidin-conjugated phycoerythrin, exhibits a 17 fold faster cleavage time than DDDK
-
-
?
SGERMMG-SA-PE + H2O
SGERMMG + SA-PE
show the reaction diagram
-
peptide conjugated to fluorescent probe streptavidin-conjugated phycoerythrin
-
-
?
thiobenzyl benzyloxy-carbonyl-L-lysinate + H2O
3-carboxy-4-nitrophenoxide + ?
show the reaction diagram
-
-
-
-
?
thiobenzyl benzyloxycarbonyl-L-lysinate + H2O
3-carboxy-4-nitrothiophenoxide + ?
show the reaction diagram
-
Z-Lys-SBzl
-
-
?
thioredoxin-human epidermal growth factor fusion protein + H2O
thioredoxin + human epidermal growth factor
show the reaction diagram
-
Trx/hEGF
-
-
?
thioredoxin-tumor necrosis factor-related apoptosis-inducing ligand fusion protein + H2O
thioredoxin + tumor necrosis factor-related apoptosis-inducing ligand
show the reaction diagram
-
trx/TRAIL
enteropeptidase is used to cleave TRAIL from thioredoxin at enzyme-substrate molar ratio of 1 : 30,000
-
?
Tosyl-Arg methyl ester + H2O
Tosyl-Arg + methanol
show the reaction diagram
-
-
-
-
?
tosyl-Lys methyl ester + H2O
tosyl-Lys + methanol
show the reaction diagram
-
-
-
-
?
Trypsinogen + H2O
?
show the reaction diagram
trypsinogen + H2O
trypsin
show the reaction diagram
-
-
-
-
?
trypsinogen + H2O
trypsin + ?
show the reaction diagram
VDYRFL-SA-PE + H2O
VDYRFL + SA-PE
show the reaction diagram
-
peptide conjugated to fluorescent probe streptavidin-conjugated phycoerythrin
-
-
?
VLDRWM-SA-PE + H2O
VLDRWM + SA-PE
show the reaction diagram
-
peptide conjugated to fluorescent probe streptavidin-conjugated phycoerythrin
-
-
?
VRDYRM-SA-PE + H2O
VRDYRM + SA-PE
show the reaction diagram
-
peptide conjugated to fluorescent probe streptavidin-conjugated phycoerythrin
-
-
?
Z-Ala-Ala-Arg-p-nitroanilide + H2O
p-nitroaniline + Z-Ala-Ala-Arg
show the reaction diagram
-
-
-
-
?
Z-Ala-Leu-Arg-p-nitroanilide + H2O
p-nitroaniline + Z-Ala-Leu-Arg
show the reaction diagram
-
-
-
-
?
Z-Ala-Met-Arg-p-nitroanilide + H2O
p-nitroaniline + Z-Ala-Met-Arg
show the reaction diagram
-
-
-
-
?
Z-Ala-Phe-Arg-p-nitroanilide + H2O
p-nitroaniline + Z-Ala-Phe-Arg
show the reaction diagram
-
-
-
-
?
Z-Ala-Trp-Arg-p-nitroanilide + H2O
Z-Ala-Trp-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Z-Ala-Tyr-Arg-p-nitroanilide + H2O
p-nitroaniline + Z-Ala-Tyr-Arg
show the reaction diagram
-
-
-
-
?
Z-Gly-Pro-Arg-p-nitroanilide + H2O
p-nitroaniline + Z-Gly-Pro-Arg
show the reaction diagram
-
-
-
-
?
Z-Pro-Phe-Arg-p-nitroanilide + H2O
p-nitroaniline + Z-Pro-Phe-Arg
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
trypsinogen + H2O
trypsin
show the reaction diagram
activation by by endoproteolytic cleavage
-
-
?
trypsinogen + H2O
trypsin + ?
show the reaction diagram
protein C inhibitor + H2O
?
show the reaction diagram
-
-
-
-
?
Trypsinogen + H2O
?
show the reaction diagram
trypsinogen + H2O
trypsin + ?
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
CaCl2 enhances enzyme activity at 0.02 mM
K+
-
activating, thiobenzyl benzyloxycarbonyl-L-lysinate as non-specific substrate
Na+
-
activating, thiobenzyl benzyloxycarbonyl-L-lysinate as non-specific substrate
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
basic pancreatic trypsin inhibitor
reversible inhibition
-
Soybean trypsin inhibitor
-
-
antipain
-
85-95% loss of activtity
Aprotinin
-
40-90% loss of activtity
benzamidine
-
75-92% loss of activtity
bestatin
-
7% loss of activtity
chymostatin
-
20% loss of activtity
dithiothreitol
-
10% loss of activity, thiobenzyl benzyloxycarbonyl-L-lysinate as substrate
E-64
-
50-85% loss of activtity
ethanol
-
80-40% loss of activity, thiobenzyl benzyloxycarbonyl-L-lysinate as substrate
imidazole
-
90-30% loss of activity, thiobenzyl benzyloxycarbonyl-L-lysinate as substrate
leupeptin
-
94% loss of activtity
N-tosyl-L-lysine chloromethyl ketone
-
13% loss of activtity
NaCl
-
at 100 and 250 mM, Gly-Asp-Asp-Asp-Asp-Lys-?-naphthylamide as substrate
pepstatin
-
5% loss of activtity
pepstatin A
-
5% loss of activtity
PMSF
-
50% loss of activtity
Protein C inhibitor
-
low molecular weight and unfractionated heparin slightly reduce the inhibitory effect of protein C inhibitor
-
SDS
-
40-70% loss of activity, thiobenzyl benzyloxycarbonyl-L-lysinate as substrate
Soybean trypsin inhibitor
-
Triton X-100
-
30-80% loss of activity, thiobenzyl benzyloxycarbonyl-L-lysinate as substrate
Tween-20
-
20-75% loss of activity, thiobenzyl benzyloxycarbonyl-L-lysinate as substrate
Urea
-
50-70% loss of activity, thiobenzyl benzyloxycarbonyl-L-lysinate as substrate
additional information
-
EDTA not inhibitory
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00258 - 0.018
GD4R-4-nitroanilide
0.0179 - 0.276
GDDDDK-4-nitroanilide
0.123 - 0.16
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
0.108
Gly-L-Asp-L-Asp-L-Asp-L-Asp-L-Lys-2-naphthylamide
at pH 8.0 and 37°C
0.14
thiobenzyl benzyloxy-carbonyl-L-lysinate
pH 8.4, 37°C
0.165 - 0.19
Z-Lys-SBzl
0.0021
A4-mutant trypsinogen
-
37 °C, 100 mM Tris-HCl (pH 8.0), 1 mM CaCl2, 120 nM soybean trypsin inhibitor, 0.13 nM human enteropeptidase
-
0.0647
APFDDDDKIVGG
-
EP light chain
0.076
formyl-Ala-Phe-Arg-4-nitroanilide
-
pH 8.4, 25°C
0.167
formyl-Ala-Phe-Lys-4-nitroanilide
-
pH 8.4, 25°C
0.5 - 0.6
GD4K 2-naphthylamide
-
pH 8.4, 35°C
0.025
GD4K 7-amido-4-methylcoumarin
-
pH 8.4, 35°C
0.16
GD4KF(NO2)G
-
50 mM Ca2+, pH 8.0, 37°C
0.019 - 0.02
GDDDDK-2-naphthylamide
0.034
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
pH 8.4, 37°C
0.034 - 0.08
Gly-Asp-Asp-Asp-Asp-Lys-naphthylamide
0.2 - 0.525
glycyl-tetra-L-aspartyl-L-lysine beta-naphthylamide
0.0014 - 0.0072
human cationic trypsinogen
-
596
N-alpha-benzyloxycarbonyl-L-lysine thiobenzyl ester +
-
37°C, pH 8.0
-
0.076
N-formyl-Ala-Phe-Arg-4-nitroanilide
-
-
0.167
N-formyl-Ala-Phe-Lys-4-nitroanilide
-
-
0.14
thiobenzyl benzyloxy-carbonyl-L-lysinate
-
pH 8.4, 25°C
0.14
thiobenzyl benzyloxycarbonyl-L-lysinate
-
-
0.0072
Trypsinogen
-
human trypsinogen
0.356
Z-Ala-Ala-Arg-nitroanilide
-
-
0.234
Z-Ala-Leu-Arg-nitroanilide
-
-
0.42
Z-Ala-Met-Arg-nitroanilide
-
-
0.067
Z-Ala-Phe-Arg-nitroanilide
-
-
0.24
Z-Ala-Trp-Arg-nitroanilide
-
-
0.109
Z-Ala-Tyr-Arg-nitroanilide
-
-
0.091
Z-Gly-Pro-Arg-nitroanilide
-
-
0.265
Z-Pro-Phe-Arg-nitroanilide
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
42 - 49
GD4R-4-nitroanilide
122 - 148
GDDDDK-4-nitroanilide
98 - 121
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
58.38
Gly-L-Asp-L-Asp-L-Asp-L-Asp-L-Lys-2-naphthylamide
at pH 8.0 and 37°C
133
thiobenzyl benzyloxy-carbonyl-L-lysinate
pH 8.4, 37°C
319 - 354
Z-Lys-SBzl
11.2
A4-mutant trypsinogen
-
37 °C, 100 mM Tris-HCl (pH 8.0), 1 mM CaCl2, 120 nM soybean trypsin inhibitor, 0.13 nM enteropeptidase
-
23.5
formyl-Ala-Phe-Arg-4-nitroanilide
-
pH 8.4, 25°C
18.2
formyl-Ala-Phe-Lys-4-nitroanilide
-
pH 8.4, 25°C
0.97 - 17.33
G5DKF(NO2)G
24.9
GD4K 2-naphthylamide
-
pH 8.4, 35°C
64.8
GD4K 7-amido-4-methylcoumarin
-
pH 8.4, 35°C
0.99 - 17.83
GD4KF(NO2)G
92 - 115
GDDDDK-2-naphthylamide
118
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
pH 8.4, 37°C
0.97 - 21.5
glycyl-tetra-L-aspartyl-L-lysine beta-naphthylamide
2.3 - 35.1
human cationic trypsinogen
-
0.07
LTAEEKA
-
pH 8.0, 37°C
111
N-alpha-benzyloxycarbonyl-L-lysine thiobenzyl ester
-
37°C, pH 8.0
133
thiobenzyl benzyloxy-carbonyl-L-lysinate
-
pH 8.4, 25°C
2.8
Trypsinogen
-
human trypsinogen
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2330 - 18900
GD4R-4-nitroanilide
537 - 6830
GDDDDK-4-nitroanilide
805 - 846
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
540.56
Gly-L-Asp-L-Asp-L-Asp-L-Asp-L-Lys-2-naphthylamide
at pH 8.0 and 37°C
1770 - 2150
Z-Lys-SBzl
309
formyl-Ala-Phe-Arg-4-nitroanilide
-
pH 8.4, 25°C
109
formyl-Ala-Phe-Lys-4-nitroanilide
-
pH 8.4, 25°C
42
GD4K 2-naphthylamide
-
pH 8.4, 35°C
2623
GD4K 7-amido-4-methylcoumarin
-
pH 8.4, 35°C
4600 - 6052
GDDDDK-2-naphthylamide
3500
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
pH 8.4, 37°C
950
thiobenzyl benzyloxy-carbonyl-L-lysinate
-
pH 8.4, 25°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0000461 - 0.000228
basic pancreatic trypsin inhibitor
-
0.0000023
Soybean trypsin inhibitor
pH 8.4, 37°C
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2100
-
thioredoxin/human epidermal growth factor fusion protein as substrate
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.4
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 9
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
assay at
25
-
assay at, substrate thiobenzyl benzyloxy-carbonyl-L-lysinate
37
-
assay at, substrate Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
enterokinase is expressed in the proximal part, the heavy chain anchors the enzyme in the intestinal brush border membrane
Manually annotated by BRENDA team
-
jejunal mucosa
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
transmembrane protein
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
physiological function
additional information
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
ENTK_HUMAN
1019
1
112935
Swiss-Prot
Secretory Pathway (Reliability: 2)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26000
x * 26000, recombinant catalytic subunit, SDS-PAGE
27000
x * 27000, deglycosylated recombinant enzyme, SDS-PAGE, x * 37000-72000, hyperglycosylated recombinant enzyme, SDS-PAGE
240000
-
x * 240000 + x * 260000, SDS-PAGE
260000
-
x * 240000 + x * 260000, SDS-PAGE
296000
-
gel filtration
30000
-
x * 30000, enteropeptidase catalytic subunit, SDS-PAGE
50000
-
x * 50000, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterodimer
-
-
trimer
-
-
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
-
secreted as a proenzyme, enterokinase converts trypsinogen to trypsin
side-chain modification
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, using 20% (w/v) polyethylene glycol 10000, 0.1 M HEPES, pH 7.5
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C112S
site-directed mutagenesis, replacement of the free cysteine residue with serine improves the refolding yield of the recombinant enzyme by 50%. The heat stability of this C112S variant was also significantly improved by supercharging
N6D/G21D/G22D/N141D/K209E
site-directed mutagenesis, the mutations lead to supercharging of the protein surface leading to 100fold increased protein solubility
N6D/G21D/G22D/N142D/K210E/C112S
supercharged variant with increased solubility more than 100fold, used for crystallization
R96Q
the mutant shows decreased specificities for substrates containing the sequences DDDDK and DDDDR, while basic pancreatic trypsin inhibitor inhibition is increased
Y174R
the mutant shows improved specificities for substrates containing the sequences DDDDK and DDDDR, while basic pancreatic trypsin inhibitor inhibition is significantly decreased
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
60
purified recombinant enteropeptidase light chain wild-type and N6D/G21D/G22D/N141D/K209E mutant enteropeptidase light chains show 60% remaining activity
65
purified recombinant enteropeptidase light chain wild-type and mutant C112S show 10% remaining activity, the purified recombinant N6D/G21D/G22D/N141D/K209E mutant enteropeptidase light chain shows 20% activity remaining
70
purified recombinant enteropeptidase light chain wild-type and mutant C112S are inactivated, purified recombinant N6D/G21D/G22D/N141D/K209E mutant enteropeptidase light chain shows 20% activity remaining
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
highly resistant to detergent to triton X-100, tween 20 and SDS
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
100% activity during storage at –20°C for at least for 1.5 years
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
amylose affinity column chromatography
HiTrap Q column chromatography
refolded, soluble, and detagged recombinant enteropeptidase light chain by affinity chromatography on soybean trypsin inhibitor agarose
soybean trypsin inhibitor-agarose column chromatography, and gel filtration
STI-agarose column chromatography and Superdex 200 gel filtration
solubilization of inclusion bodies with 6 M guanidine-HCl, affinity chromatography on STI-agarose
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells with maltose binding protein tag
expressed in Escherichia coli BL21(DE3) codon plus RIL hEPL sc C122S cells
expressed in Pichia pastoris
expressed in Pichia pastoris strain Y11430
expressed in Pichia pastoris X-33 strain
expression of light chain of enteropeptidase in Escherichia coli BL21(DE3) in both soluble and insoluble form
recombinant expression of enteropeptidase light chain as thioredoxin-fusion protein in Escherichia coli strain BL21(DE3) in aggregated form, subcloning in Escherichia coli strain DH5alpha
the synthetic gene encoding human enteropeptidase light chain with His-tag added at the C-terminus to facilitate protein purification is cloned into Pichia pastoris expression plasmids under the control of an inducible AOX1 or constitutive promoters GAP and AAC
expressed in Escherichia coli strain BL21(DE3) and on M13 filamentous bacteriophage
-
expression as fusion protein with thioredoxin in Escherichia coli
-
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
fast dilution method in 0.7 M arginine, 1 mM EDTA, 3 mM reduced glutathione, and 3 mM GSSG at pH 8.6
recombinant expression of thioredoxin-fusion enteropeptidase light chain from aggregates in 3 M guanidine-HCl, pH 8.0, in Escherichia coli strain BL21(DE3) by dilution of the protein in 0.7 M arginine-HCl, pH 8.5, 15% v/v glycerol, 3 mM reduced glutathione, and 1 mM EDTA. After 72 h at 4°C the refolding solution is dialyzed for 8 h against 50 mM Tris-HCl, pH 8.0, to facilitate complete autocatalytic activation by cleaving the fusion tag
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
enteropeptidase is a serine protease used in different biotechnological applications. For many applications the smaller light chain can be used to avoid the expression of the rather large holoenzyme
molecular biology
the high specificity of the target site makes enterokinase an ideal tool for cleaving fusion proteins at defined cleavage sites
synthesis
the enzyme can be used for cleavage of fusion proteins due to its high specific activity
analysis
-
cellular libraries of peptide substrates, CLiPS, are used to study substrate specificities, fluorescent reporter substrates on the surface of Escherichia coli as N-terminal conjugates are used as whole-cell protease activity assays
medicine
-
human TRAIL is a candidate for clinical application in cancer therapy, activity is lost in some forms of recombinant TRAIL, refolding of thioredoxin/TRAIL and cleavage by enteropeptidase yield a biological active anticancer agent
molecular biology
-
the high degree of specificity exhibited by enteropeptidase makes it a suitable reagent for cleaving recombinant proteins to remove affinity or other tags. However often unwanted cleavages elsewhere in the protein occur during cleavage of fusions when high amount of enzyme is required
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Grant, D.A.W.; Hermon-Taylor, J.
Enterokinase
Methods Enzym. Anal. , 3rd Ed. (Bergmeyer, H. U. , ed. )
5
143-155
1984
Bos taurus, Homo sapiens, Sus scrofa
-
Manually annotated by BRENDA team
Magee, A.I.; Grant, D.A.W.; Hermon-Taylor, J.; Offord, R.E.
Specific one-stage method for assay of enterokinase activity by release of radiolabelled activation peptides from alpha-N-[3H]acetyl-trypsinogen and the effect of calcium ions on the enzyme activity
Biochem. J.
197
239-244
1981
Homo sapiens
Manually annotated by BRENDA team
Brodrick, J.W.; Largman, C.; Hsiang, M.W.; Johnson, J.H.; Geokas, M.C.
Structural basis for the specific activation of human enteropeptidase
J. Biol. Chem.
253
2737-2742
1978
Homo sapiens, Sus scrofa
Manually annotated by BRENDA team
Grant, D.A.W.; Hermon-Taylor, J.
The purification of human enterokinase by affinity chromatography and immunoadsorption. Some observations on its molecular characteristics and comparisons with the pig enzyme
Biochem. J.
155
243-254
1976
Homo sapiens, Sus scrofa
Manually annotated by BRENDA team
Miyoshi, Y.; Onishi, T.; Sano, T.; Komi, N.
Monoclonal antibody against human enterokinase and immunohistochemica localization of the enzyme
Gastroenterol. Jpn.
25
320-327
1990
Homo sapiens
Manually annotated by BRENDA team
Mann, N.S.; Mann, S.K.
Enterokinase
Proc. Soc. Exp. Biol. Med.
206
114-118
1994
Homo sapiens
Manually annotated by BRENDA team
Light, A.; Janska, G.
Enterokinase (enteropeptidase): comparative aspects
Trends Biochem. Sci.
14
110-112
1989
Bos taurus, Homo sapiens, Sus scrofa
Manually annotated by BRENDA team
Gasparian, M.E.; Ostapchenko, V.G.; Schulga, A.A.; Dolgikh, D.A.; Kirpichnikov, M.P.
Expression, purification, and characterization of human enteropeptidase catalytic subunit in Escherichia coli
Protein Expr. Purif.
31
133-139
2003
Homo sapiens (P98073), Homo sapiens
Manually annotated by BRENDA team
Mikhailova, A.G.; Likhareva, V.V.; Prudchenko, I.A.; Rumsh, L.D.
Effect of calcium ions on enteropeptidase catalysis
Biochemistry (Moscow)
70
1129-1135
2005
Homo sapiens
Manually annotated by BRENDA team
Nemoda, Z.; Sahin-Toth, M.
The tetra-aspartate motif in the activation peptide of human cationic trypsinogen is essential for autoactivation control but not for enteropeptidase recognition
J. Biol. Chem.
280
29645-29652
2005
Bos taurus, Homo sapiens
Manually annotated by BRENDA team
Gasparian, M.E.; Ostapchenko, V.G.; Dolgikh, D.A.; Kirpichnikov, M.P.
Biochemical characterization of human enteropeptidase light chain
Biochemistry (Moscow)
71
113-119
2006
Bos taurus, Homo sapiens
Manually annotated by BRENDA team
Gasparian, M.E.; Ostapchenko, V.G.; Yagolovich, A.V.; Tsygannik, I.N.; Chernyak, B.V.; Dolgikh, D.A.; Kirpichnikov, M.P.
Overexpression and refolding of thioredoxin/TRAIL fusion from inclusion bodies and further purification of TRAIL after cleavage by enteropeptidase
Biotechnol. Lett.
29
1567-1573
2007
Homo sapiens
Manually annotated by BRENDA team
Boulware, K.T.; Daugherty, P.S.
Protease specificity determination by using cellular libraries of peptide substrates (CLiPS)
Proc. Natl. Acad. Sci. USA
103
7583-7588
2006
Homo sapiens
Manually annotated by BRENDA team
Mikhailova, A.G.; Likhareva, V.V.; Teich, N.; Rumsh, L.D.
The ways of realization of high specificity and efficiency of enteropeptidase
Protein Pept. Lett.
14
227-232
2007
Homo sapiens, Bos taurus (Q6B4R4), Bos taurus
Manually annotated by BRENDA team
Makarova, A.M.; Gorbacheva, L.R.; Savinkova, I.V.; Mikhailova, A.G.; Rumsh, L.D.; Pinelis, V.G.; Strukova, S.M.
Effect of enteropeptidase on survival of cultured hippocampal neurons under conditions of glutamate toxicity
Biochemistry (Moscow)
75
1153-1159
2010
Bos taurus, Homo sapiens
Manually annotated by BRENDA team
Choi, M.G.; Lee, E.; Chung, H.S.; Jang, S.H.; Lee, C.
A fluorogenic method for measuring enteropeptidase activity: spectral shift in the emission of GD4K-conjugated 7-amino-4-methylcoumarin
BMB Rep.
44
458-461
2011
Homo sapiens
Manually annotated by BRENDA team
Pepeliaev, S.; Krahulec, J.; ?erny, Z.; Jilkova, J.; Tlusta, M.; Dostalova, J.
High level expression of human enteropeptidase light chain in Pichia pastoris
J. Biotechnol.
156
67-75
2011
Homo sapiens (P98073), Homo sapiens
Manually annotated by BRENDA team
Simeonov, P.; Berger-Hoffmann, R.; Hoffmann, R.; Straeter, N.; Zuchner, T.
Surface supercharged human enteropeptidase light chain shows improved solubility and refolding yield
Protein Eng. Des. Sel.
24
261-268
2011
Homo sapiens (P98073), Homo sapiens
Manually annotated by BRENDA team
Gasparian, M.E.; Bychkov, M.L.; Dolgikh, D.A.; Kirpichnikov, M.P.
Strategy for improvement of enteropeptidase efficiency in tag removal processes
Protein Expr. Purif.
79
191-196
2011
Bos taurus, Homo sapiens
Manually annotated by BRENDA team
Gasparian, M.E.; Bobik, T.V.; Kim, Y.V.; Ponomarenko, N.A.; Dolgikh, D.A.; Gabibov, A.G.; Kirpichnikov, M.P.
Heterogeneous catalysis on the phage surface: Display of active human enteropeptidase
Biochimie
95
2076-2081
2013
Homo sapiens
Manually annotated by BRENDA team
Ostapchenko, V.G.; Gasparian, M.E.; Kosinsky, Y.A.; Efremov, R.G.; Dolgikh, D.A.; Kirpichnikov, M.P.
Dissecting structural basis of the unique substrate selectivity of human enteropeptidase catalytic subunit
J. Biomol. Struct. Dyn.
30
62-73
2012
Bos taurus, Homo sapiens
Manually annotated by BRENDA team
Jirickova, K.; Gal, M.; Krahulec, J.; Hives, J.
Electrochemical determination of basic biochemical properties of enzyme enterokinase
Monatsh. Chem.
146
755-759
2015
Homo sapiens (P98073)
-
Manually annotated by BRENDA team
Prohaska, T.A.; Wahlmueller, F.C.; Furtmueller, M.; Geiger, M.
Interaction of protein C inhibitor with the type II transmembrane serine protease enteropeptidase
PLoS ONE
7
e39262
2012
Homo sapiens
Manually annotated by BRENDA team
Smith, E.T.; Johnson, D.A.
Human enteropeptidase light chain: bioengineering of recombinants and kinetic investigations of structure and function
Protein Sci.
22
577-585
2013
Homo sapiens (P98073), Homo sapiens
Manually annotated by BRENDA team
Simeonov, P.; Zahn, M.; Straeter, N.; Zuchner, T.
Crystal structure of a supercharged variant of the human enteropeptidase light chain
Proteins
80
1907-1910
2012
Homo sapiens (P98073), Homo sapiens
Manually annotated by BRENDA team
Melicherova, K.; Krahulec, J.; Safranek, M.; Liskova, V.; Hopkova, D.; Szeliova, D.; Turna, J.
Optimization of the fermentation and downstream processes for human enterokinase production in Pichia pastoris
Appl. Microbiol. Biotechnol.
101
1927-1934
2017
Homo sapiens (P98073), Homo sapiens
Manually annotated by BRENDA team
Niu, L.; Li, J.; Ji, X.; Yang, B.
Efficient expression and purification of recombinant human enteropeptidase light chain in Escherichia coli
Braz. Arch. Biol. Technol.
58
216-221
2015
Homo sapiens (P98073)
-
Manually annotated by BRENDA team
Hayashi, H.; Kubo, Y.; Izumida, M.; Takahashi, E.; Kido, H.; Sato, K.; Yamaya, M.; Nishimura, H.; Nakayama, K.; Matsuyama, T.
Enterokinase enhances influenza A virus infection by activating trypsinogen in human cell lines
Front. Cell. Infect. Microbiol.
8
91
2018
Homo sapiens (P98073), Homo sapiens
Manually annotated by BRENDA team