Information on EC 3.4.21.9 - enteropeptidase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Euteleostomi

EC NUMBER
COMMENTARY
3.4.21.9
-
RECOMMENDED NAME
GeneOntology No.
enteropeptidase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Activation of trypsinogen by selective cleavage of Lys6-/-Ile bond
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
hydrolysis of peptide bond
-
-
hydrolysis of peptide bond
-
-
hydrolysis of peptide bond
A4UWM5, -
-
hydrolysis of peptide bond
-
-
endopeptidase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
BEP
-
-
bovine enterokinase light chain
-
recombinant
bovine enteropeptidase
-
-
Chinese bovine enterokinase
-
catalytic subunit
Chinese northern yellow bovine enterokinase catalytic subunit
-
-
EC 3.4.4.8
-
-
formerly
-
EKL
-
-
EKL-His6
-
-
enterokinase
-
-
-
-
enterokinase
Q6B4R4
EK
enterokinase
-
-
enterokinase
P98073
-
enterokinase
-
native
enterokinase
synthetic construct
-
-
enterokinase light chain
-
recombinant
enteropeptidase
-
-
enteropeptidase
-
EK
enteropeptidase
Q6B4R4
EP
enteropeptidase
-
-
enteropeptidase
-
EP light chain
enteropeptidase
A4UWM5
EP
enteropeptidase light chain
-
L-BEP
enteropeptidase light chain
-
L-HEP
EP 118-1035
-
-
EP-1
A4UWM5
-
human enteropeptidase
-
-
native enterokinase
-
-
natural enteropeptidase
-
-
peptidase, entero-
-
-
-
-
porcine enterokinase
-
-
recombinant bovine enterokinase catalytic subunit protein
-
-
recombinant enterokinase light chain
-
-
recombinant His-tagged enterokinase light chain
-
-
rEKL/His
-
-
sBEKLC
-
gene
CAS REGISTRY NUMBER
COMMENTARY
9014-74-8
-
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
enteropeptidase is a serine protease secreted by the pancreas that converts inactive trypsinogen to active trypsin
physiological function
-
the enzyme converts the inactive form of trypsinogen to active trypsin by endoproteolytic cleavage
additional information
-
low enzyme concentrations protect hippocampal neurons against the death caused by 0.1 mM glutamate, via a proteinase activated receptor PAR1-dependent mechanism, while High concentrations of the enzyme causes the death of neurons mainly through necrosis
additional information
-
low enzyme light chain concentrations protect hippocampal neurons against the death caused by 0.1 mM glutamate, via a proteinase activated receptor PAR1-dependent mechanism, while high concentrations of enzyme light chain causes the death of neurons mainly through necrosis
additional information
-
enterokinase shows no strict cleavage site specificity because of sporadic cleavage at other sites, which consist of acidic and basic residues
additional information
-
improved efficiency of fusion proteins cleavage by enteropeptidase by substitution of the Lys residue by Arg in specific cleavage sequence (Asp)4-Lys. 3-6fold lower amounts of the catalytic subunit of enteropeptidase is required for 95% cleavage of Trx/TRAIL and Trx/FGF-2 fusions with (Asp)4-Arg cleavage sequence in comparison to native sequence (Asp)4-Lys. Reduced amount of non-specifically cleaved peptide fragments are observed during cleavage of (Asp)4-Lys/Arg mutated fusions
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
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
-
-
?
AhR6-C/EBP
?
show the reaction diagram
synthetic construct
-
hybrid of the AhR basic region and C/EBP leucine zipper. Reaction under standard conditions (at 37C in 50 mM Tris buffer, pH 7.6) is rapid and somewhat nonspecific. After just 15 min incubation, 82% of the protein has been cleaved at multiple sites, prolonged incubation causes further degradation of cleavage products
-
-
?
alpha-benzoyl-DL-Arg-4-nitroanilide + H2O
alpha-benzoyl-DL-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Angiotensin II + H2O
?
show the reaction diagram
-
-
-
-
?
APFDDDDKIVGG + H2O
?
show the reaction diagram
-
N-terminal dodecapeptides of human cationic trypsinogen
-
-
?
APFDDDDKIVGG + H2O
?
show the reaction diagram
Q6B4R4
N-terminal dodecapeptides of human cationic trypsinogen
-
-
?
APFDDDGKIVGG + H2O
?
show the reaction diagram
Q6B4R4
N-terminal dodecapeptides of human pancreatitis-associated mutant variant of trypsinogen
-
-
?
APFDDDGRIVGG + H2O
?
show the reaction diagram
Q6B4R4
N-terminal dodecapeptides of human cationic tryosinogen
-
-
?
benzoyl-Arg ethyl ester + H2O
benzoyl-Arg + ethanol
show the reaction diagram
-
-
-
-
?
benzoyl-Arg ethyl ester + H2O
benzoyl-Arg + ethanol
show the reaction diagram
-
-
-
-
?
benzoyl-Arg ethyl ester + H2O
benzoyl-Arg + ethanol
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
-
-
?
benzyloxycarbonyl-Phe-Arg-4-methylcoumaryl 7-amide + H2O
?
show the reaction diagram
-
-
-
-
?
Boc-Glu(OBzl)-Ala-Arg-MCA + H2O
7-amino-4-methylcoumarin + Boc-Glu(OBzl)-Ala-Arg
show the reaction diagram
A4UWM5, -
-
-
-
?
bovine trypsinogen + H2O
?
show the reaction diagram
-
-
-
-
?
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
-
-
?
egg white lysozyme + H2O
?
show the reaction diagram
-
egg white lysozyme from hen
-
-
?
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
-
-
?
fusion protein Trx/hEGF + H2O
?
show the reaction diagram
-, P98073
the fusion protein Trx/hEGF contains an inter-domain enteropeptidase recognition site
-
-
?
fusion protein Trx/hEGF + H2O
?
show the reaction diagram
-, P98073
the fusion protein Trx/hEGF contains an inter-domain enteropeptidase recognition site
-
?
fusion protein Trx/hIL-13 + H2O
?
show the reaction diagram
-, P98073
the fusion protein Trx/hIL-13 contains an inter-domain enteropeptidase recognition site
-
-
?
fusion protein Trx/hIL-13 + H2O
?
show the reaction diagram
-, P98073
the fusion protein Trx/hIL-13 contains an inter-domain enteropeptidase recognition site
-
?
G5DKF(NO2)G + H2O
?
show the reaction diagram
-
-
-
-
-
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
-
-
-
-
?
GD4KF(NO2)G + H2O
?
show the reaction diagram
-
-
-
-
?
glutathione S-transferase-enterokinase + H2O
enterokinase + glutathione S-transferase
show the reaction diagram
-, Q6B4R4
GST-EK harboring an EK site
-
-
?
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide + H2O
Gly-Asp-Asp-Asp-Asp-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-Asp-Asp-Asp-Asp-Lys-2-naphthylamide + H2O
Gly-Asp-Asp-Asp-Asp-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-Asp-Asp-Asp-Asp-Lys-2-naphthylamide + H2O
?
show the reaction diagram
-
-
-
?
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide + H2O
?
show the reaction diagram
-, P98073
-
-
-
?
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide + H2O
?
show the reaction diagram
-, P98073
-
-
?
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide + H2O
2-naphthylamine + Gly-Asp-Asp-Asp-Asp-Lys
show the reaction diagram
-
-
-
-
?
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide + H2O
2-naphthylamine + Gly-Asp-Asp-Asp-Asp-Lys
show the reaction diagram
A4UWM5, -
-
-
-
?
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide + H2O
Gly-Asp-Asp-Asp-Asp + Lys-2-naphthylamide
show the reaction diagram
-
-
-
-
?
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide + H2O
Gly-Asp-Asp-Asp-Asp + Lys-2-naphthylamide
show the reaction diagram
-
-
-
-
?
Gly-Asp-Asp-Asp-Asp-Lys-beta-naphthylamide + H2O
Gly-Asp-Asp-Asp-Asp-Lys + beta-naphthylamine
show the reaction diagram
-
-
-
-
?
Gly-Asp-Asp-Asp-Asp-Lys-beta-naphthylamide + H2O
naphthylamine + Gly-Asp-Asp-Asp-Asp-Lys
show the reaction diagram
-
GD4K-na
-
-
?
Gly-Asp-Asp-Asp-Asp-Lys-Ile-Val-Gly-Gly + H2O
Gly-Asp-Asp-Asp-Asp + Lys-Ile-Val-Gly-Gly
show the reaction diagram
-
-
-
-
?
Gly-Asp-Asp-Asp-Asp-Lys-naphthylamide + H2O
naphthylamine + Gly-Asp-Asp-Asp-Asp-Lys
show the reaction diagram
-
GD4K-na
-
-
?
Gly-Gly-Asp-Asp-Asp-Lys-Ile-Val-Gly-Gly + H2O
Gly-Gly-Asp-Asp-Asp + Lys-Ile-Val-Gly-Gly
show the reaction diagram
-
-
-
-
?
Gly-Gly-Gly-Asp-Asp-Lys-Ile-Val-Gly-Gly + H2O
Gly-Gly-Gly-Asp-Asp + Lys-Ile-Val-Gly-Gly
show the reaction diagram
-
-
-
-
?
Gly-Gly-Gly-Gly-Asp-Lys-Ile-Val-Gly-Gly + H2O
Gly-Gly-Gly-Gly-Asp + Lys-Ile-Val-Gly-Gly
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
-
-
-
-
?
GST-GFPuv + H2O
GST + GFPuv
show the reaction diagram
-, Q6B4R4
harboring an EK site between GST and the green fluorescent protein GFPuv
-
-
?
GST-vasostatin fusion protein + H2O
?
show the reaction diagram
-
-
-
-
?
human cationic trypsinogen + H2O
?
show the reaction diagram
-
-
-
-
?
human thioredoxin fused human NT-proCNP(1-50) + H2O
cleavage products
show the reaction diagram
-
22kDA substrate
21 and 16 kDa cleavage products using the recombinant enzyme, multiple cleavage products using the native enzyme
-
?
interferon-alpha2a + H2O
?
show the reaction diagram
-
recombinantly expressed substrate
-
-
?
interferon-alpha2b + H2O
?
show the reaction diagram
-
recombinantly expressed substrate
-
-
?
LTAEEKA + H2O
?
show the reaction diagram
-
-
-
-
?
LTAEEKA + H2O
?
show the reaction diagram
-
Hb 2-8
-
-
?
LTAEEKAAV + H2O
?
show the reaction diagram
-
-
-
-
?
Lys-2-naphthylamide + H2O
Lys + 2-naphthylamine
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
-
-
?
MUC1-IgG Fc + H2O
MUC1 + IgG Fc
show the reaction diagram
-
fusion protein
-
-
?
mucin 1-IgG2a Fc + H2O
mucin 1 + ?
show the reaction diagram
-
cleavage of the mucin fusion protein
-
-
?
N-alpha-benzyloxycarbonyl-L-lysine thiobenzyl ester + H2O
N-alpha-benzyloxycarbonyl-L-lysine + benzylmercaptane
show the reaction diagram
-
-
-
-
?
Nalpha-acetyltrypsinogen + H2O
Nalpha-acetylactivation peptides + trypsin
show the reaction diagram
-
-
-
?
PrAD4KP26 + H2O
?
show the reaction diagram
-
fusion protein containing a modified protein A as a carrier and recoverin as a target protein
-
-
?
Pro-Phe-Arg-4-methylcoumaryl-7-amide + H2O
7-amino-4-methylcoumarin + Pro-Phe-Arg
show the reaction diagram
A4UWM5, -
-
-
-
?
S-alkylated soybean trypsin inhibitor + H2O
?
show the reaction diagram
-
limited proteolysis
-
-
?
S-carboxyamidomethyl derivative of bovine serum albumin + H2O
?
show the reaction diagram
-
bovine serum albumin is resistant in its native state, somewhat susceptible as the S-carboxyamidomethyl derivative and highly susceptible as the S-carboxymethyl derivative
-
-
?
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
-
-
?
tert-butoxycarbonyl-Gln-Ala-Arg-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
weak activity
-
-
?
tert-butoxycarbonyl-Leu-Ser-Thr-Arg-4-methylcoumaryl 7-amide + H2O
?
show the reaction diagram
-
-
-
-
?
tert-butoxycarbonyl-Leu-Thr-Arg-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
weak activity
-
-
?
tert-butoxycarbonyl-Phe-Ser-Arg-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
weak activity
-
-
?
tert-butoxycarbonyl-Val-Leu-Lys-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
weak activity
-
-
?
thiobenzyl benzyloxy-carbonyl-L-lysinate + H2O
?
show the reaction diagram
-, P98073
-
-
-
?
thiobenzyl benzyloxy-carbonyl-L-lysinate + H2O
?
show the reaction diagram
-, P98073
-
-
?
thiobenzyl benzyloxy-carbonyl-L-lysinate + H2O
3-carboxy-4-nitrophenoxide + ?
show the reaction diagram
-
-
-
-
?
thiobenzyl benzyloxycarbonyl-L-lysinate + H2O
?
show the reaction diagram
-
-
-
-
?
thiobenzyl benzyloxycarbonyl-L-lysinate + H2O
3-carboxy-4-nitrothiophenoxide + ?
show the reaction diagram
-
Z-Lys-SBzl
-
-
?
thioredoxin-DDDD156K-mouse NT-proCNP(2-40)-DDDK201SRLLR + H2O
cleavage products
show the reaction diagram
-
22kDA substrate
21 and 16 kDa cleavage products using the recombinant enzyme, multiple cleavage products using the native enzyme
-
?
thioredoxin-DDDD156RRK-mouse NT-proCNP(2-40)-DDDK201SRLLR + H2O
cleavage products
show the reaction diagram
-
22kDA substrate
21 and 16 kDa cleavage products using the recombinant enzyme, multiple cleavage products using the native enzyme
-
?
thioredoxin-DDDD156SDK-mouse NT-proCNP(2-40)-DDDK201SRLLR + H2O
cleavage products
show the reaction diagram
-
22kDA substrate
21 and 16 kDa cleavage products using the recombinant enzyme, multiple cleavage products using the native enzyme
-
?
thioredoxin-DDDD156SK-mouse NT-proCNP(2-40)-DDDK201SRLLR + H2O
cleavage products
show the reaction diagram
-
22kDA substrate, 9 fusion proteins are tested as substrates, addition of SRLLR residues leads to an increase in activity
21 and 16 kDa cleavage products using the recombinant enzyme, multiple cleavage products using the native enzyme
-
?
thioredoxin-DDDD156SRK-mouse NT-proCNP(2-40)-DDDK201SRLLR + H2O
cleavage products
show the reaction diagram
-
22kDA substrate
21 and 16 kDa cleavage products using the recombinant enzyme, multiple cleavage products using the native enzyme
-
?
thioredoxin-DDDD156SRLK-mouse NT-proCNP(2-40)-DDDK201SRLLR + H2O
cleavage products
show the reaction diagram
-
22kDA substrate
21 and 16 kDa cleavage products using the recombinant enzyme, multiple cleavage products using the native enzyme
-
?
thioredoxin-DDDD156SRLLK-mouse NT-proCNP(2-40)-DDDK201SRLLR + H2O
cleavage products
show the reaction diagram
-
22kDA substrate
21 and 16 kDa cleavage products using the recombinant enzyme, multiple cleavage products using the native enzyme
-
?
thioredoxin-DDDD156SRLLRK-mouse NT-proCNP(2-40)-DDDK201SRLLR + H2O
cleavage products
show the reaction diagram
-
22kDA substrate, best substrate for enterokinase activity
21 and 16 kDa cleavage products using the recombinant enzyme, multiple cleavage products using the native enzyme
-
?
thioredoxin-DDDD156SSK-mouse NT-proCNP(2-40)-DDDK201SRLLR + H2O
cleavage products
show the reaction diagram
-
22kDA substrate
21 and 16 kDa cleavage products using the recombinant enzyme, multiple cleavage products using the native enzyme
-
?
thioredoxin-fused N-terminal pro-C-type natriuretic peptide + H2O
N-terminal pro-C-type natriuretic peptide fragment + ?
show the reaction diagram
-
-
-
-
?
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-Arg methyl ester + H2O
Tosyl-Arg + methanol
show the reaction diagram
-
-
-
-
?
Tosyl-Arg methyl ester + H2O
Tosyl-Arg + methanol
show the reaction diagram
-
-
-
-
?
tosyl-Lys methyl ester + H2O
tosyl-Lys + 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
?
show the reaction diagram
-
-
-
-
-
Trypsinogen + H2O
?
show the reaction diagram
-
-
-
-
-
Trypsinogen + H2O
?
show the reaction diagram
-
-
-
-
?
Trypsinogen + H2O
?
show the reaction diagram
-
initiation of digestive enzyme activation by converting trypsinogen into trypsin
-
-
-
Trypsinogen + H2O
?
show the reaction diagram
-
initiates activation of pancreatic hydrolases by cleaving and activating trypsinogen
-
-
-
Trypsinogen + H2O
?
show the reaction diagram
-
enterokinase deficiency is a distinct clinical entity characterized by diarrhea, failure to thrive, hypoproteinemia, and edema. Acquired enterokinase deficiency may occur in some diffuse small bowel diseases. Steatorhea of cellac sprue may be due partly to the fact that deficiency of secretin and cholecystokinin may interfere with the action of enterokinase
-
-
-
Trypsinogen + H2O
?
show the reaction diagram
-
the enzyme plays a key role in initiating the proteolytic digestion cascade in duodenum by converting trypsinogen to trypsin
-
-
-
Trypsinogen + H2O
?
show the reaction diagram
-
physiological activator of trypsinogen
-
-
-
Trypsinogen + H2O
?
show the reaction diagram
-
rapid activation of trypsinogen in pancreatic juice. The enzyme has a key permissive role in protein digestion and plays an essential part in the zymogen mechanism that protects the pancreas from self-destruction
-
-
-
trypsinogen + H2O
trypsin + ?
show the reaction diagram
-
-
-
?
trypsinogen + H2O
trypsin + ?
show the reaction diagram
-
-
-
-
?
trypsinogen + H2O
trypsin + ?
show the reaction diagram
-
-
-
?
trypsinogen + H2O
trypsin + ?
show the reaction diagram
-
-
-
-
?
trypsinogen + H2O
trypsin + ?
show the reaction diagram
-
-
-
?
trypsinogen + H2O
trypsin + ?
show the reaction diagram
-
-
-
-
?
trypsinogen + H2O
trypsin + ?
show the reaction diagram
A4UWM5, -
-
-
-
?
trypsinogen + H2O
trypsin + ?
show the reaction diagram
-
human cationic trypsinogen is activated much more readily than bovine trypsinogen
-
-
?
trypsinogen + H2O
trypsin + ?
show the reaction diagram
-
specificity for the sequence (Asp)4-Lys-Ile
-
-
?
trypsinogen + H2O
trypsin + ?
show the reaction diagram
-
bovine trypsinogen
-
-
?
trypsinogen + H2O
trypsin + ?
show the reaction diagram
-
cleaved exclusively at the Lys6-Ile7-peptide bond
-
-
?
trypsinogen + H2O
trypsin + ?
show the reaction diagram
-
the specificity site of enterokinase recognizes in trypsinogen not merely the basic residue of the -Lys6-/-Ile7-bond, which is split during activation of the zymogen but also recognizes the sequence -Asp4-Lys, residues 2 to 6, which is present in all of the trypsinogens so far studied
-
-
?
trypsinogen + H2O
trypsin
show the reaction diagram
-
-
-
-
?
trypsinogen + H2O
trypsin
show the reaction diagram
-
-
-
-
?
trypsinogen + H2O
trypsin
show the reaction diagram
-, Q6B4R4
-
-
-
?
trypsinogen + H2O
trypsin
show the reaction diagram
Q6B4R4
-
-
-
?
trypsinogen + H2O
trypsin
show the reaction diagram
P98073
activation by by endoproteolytic cleavage, the enzyme cleaves after lysine residue if the Lys is preceded by four Asp and not followed by a Pro, and it shows high specificity of the target site. The light chain represents the catalytic enzyme subunit
-
-
?
Val-(Ala)2-(Asp)2-Lys-Ile-Val-Gly + H2O
Val-(Ala)2-(Asp)2-Lys + Ile-Val-Gly
show the reaction diagram
P98074
107% relative activity
-
-
?
Val-(Ala)2-(Asp)2-Lys-Ile-Val-Gly + H2O
Val-(Ala)2-(Asp)2-Lys + Ile-Val-Gly
show the reaction diagram
P98072
109.3% relative activity
-
-
?
Val-(Ala)2-Asp-Ala-Lys-Ile-Val-Gly + H2O
Val-(Ala)2-Asp-Ala-Lys + Ile-Val-Gly
show the reaction diagram
P98072
16.8% relative activity
-
-
?
Val-(Ala)2-Asp-Ala-Lys-Ile-Val-Gly + H2O
Val-(Ala)2-Asp-Ala-Lys + Ile-Val-Gly
show the reaction diagram
P98074
21.1% relative activity
-
-
?
Val-(Ala)3-Asp-Lys-Ile-Val-Gly + H2O
Val-(Ala)3-Asp-Lys + Ile-Val-Gly
show the reaction diagram
P98072
75.6% relative activity
-
-
?
Val-(Ala)3-Asp-Lys-Ile-Val-Gly + H2O
Val-(Ala)3-Asp-Lys + Ile-Val-Gly
show the reaction diagram
P98074
81.2% relative activity
-
-
?
Val-(Asp)2-Ala-Asp-Lys-Ile-Val-Gly + H2O
Val-(Asp)2-Ala-Asp-Lys + Ile-Val-Gly
show the reaction diagram
P98072
76.1% relative activity
-
-
?
Val-(Asp)2-Ala-Asp-Lys-Ile-Val-Gly + H2O
Val-(Asp)2-Ala-Asp-Lys + Ile-Val-Gly
show the reaction diagram
P98074
93.1% relative activity
-
-
?
Val-(Asp)3-Ala-Lys-Ile-Val-Gly + H2O
Val-(Asp)3-Ala-Lys + Ile-Val-Gly
show the reaction diagram
P98072
16.3% relative activity
-
-
?
Val-(Asp)3-Ala-Lys-Ile-Val-Gly + H2O
Val-(Asp)3-Ala-Lys + Ile-Val-Gly
show the reaction diagram
P98074
20.2% relative activity
-
-
?
Val-(Asp)3-Glu-Lys-Ile-Val-Gly + H2O
Val-(Asp)3-Glu-Lys + Ile-Val-Gly
show the reaction diagram
P98072
42.5% relative activity
-
-
?
Val-(Asp)3-Glu-Lys-Ile-Val-Gly + H2O
Val-(Asp)3-Glu-Lys + Ile-Val-Gly
show the reaction diagram
P98074
64.9% relative activity
-
-
?
Val-(Asp)4-Arg-Ile-Val-Gly + H2O
Val-(Asp)4-Arg + Ile-Val-Gly
show the reaction diagram
P98072
115.2% relative activity
-
-
?
Val-(Asp)4-Arg-Ile-Val-Gly + H2O
Val-(Asp)4-Arg + Ile-Val-Gly
show the reaction diagram
P98074
128.2% relative activity
-
-
?
Val-(Asp)4-Lys-Ile-Val-Gly + H2O
Val-(Asp)4-Lys + Ile-Val-Gly
show the reaction diagram
P98072
100% relative activity
-
-
?
Val-(Asp)4-Lys-Ile-Val-Gly + H2O
Val-(Asp)4-Lys + Ile-Val-Gly
show the reaction diagram
P98074
100% relative activity
-
-
?
Val-Ala-(Asp)2-Ala-Lys-Ile-Val-Gly + H2O
Val-Ala-(Asp)2-Ala-Lys + Ile-Val-Gly
show the reaction diagram
P98072
4.3% relative activity
-
-
?
Val-Ala-(Asp)2-Ala-Lys-Ile-Val-Gly + H2O
Val-Ala-(Asp)2-Ala-Lys + Ile-Val-Gly
show the reaction diagram
P98074
5.6% relative activity
-
-
?
Val-Ala-(Asp)3-Lys-Ile-Val-Gly + H2O
Val-Ala-(Asp)3-Lys + Ile-Val-Gly
show the reaction diagram
P98072
104.2% relative activity
-
-
?
Val-Ala-(Asp)3-Lys-Ile-Val-Gly + H2O
Val-Ala-(Asp)3-Lys + Ile-Val-Gly
show the reaction diagram
P98074
123.7% relative activity
-
-
?
Val-Ala-Asp-Ala-Asp-Lys-Ile-Val-Gly + H2O
Val-Ala-Asp-Ala-Asp-Lys + Ile-Val-Gly
show the reaction diagram
P98074
35.8% relative activity
-
-
?
Val-Ala-Asp-Ala-Asp-Lys-Ile-Val-Gly + H2O
Val-Ala-Asp-Ala-Asp-Lys + Ile-Val-Gly
show the reaction diagram
P98072
48.1% relative activity
-
-
?
Val-Asp-(Ala)2-Asp-Lys-Ile-Val-Gly + H2O
Val-Asp-(Ala)2-Asp-Lys + Ile-Val-Gly
show the reaction diagram
P98072
82.6% relative activity
-
-
?
Val-Asp-(Ala)2-Asp-Lys-Ile-Val-Gly + H2O
Val-Asp-(Ala)2-Asp-Lys + Ile-Val-Gly
show the reaction diagram
P98074
95.8% relative activity
-
-
?
Val-Asp-(Ala)3-Lys-Ile-Val-Gly + H2O
Val-Asp-(Ala)3-Lys + Ile-Val-Gly
show the reaction diagram
P98074
6.6% relative activity
-
-
?
Val-Asp-(Ala)3-Lys-Ile-Val-Gly + H2O
Val-Asp-(Ala)3-Lys + Ile-Val-Gly
show the reaction diagram
P98072
7.1% relative activity
-
-
?
Val-Asp-Ala-(Asp)2-Lys-Ile-Val-Gly + H2O
Val-Asp-Ala-(Asp)2-Lys + Ile-Val-Gly
show the reaction diagram
P98072
117.4% relative activity
-
-
?
Val-Asp-Ala-(Asp)2-Lys-Ile-Val-Gly + H2O
Val-Asp-Ala-(Asp)2-Lys + Ile-Val-Gly
show the reaction diagram
P98074
119.7% relative activity
-
-
?
Val-Asp-Ala-Asp-Ala-Lys-Ile-Val-Gly + H2O
Val-Asp-Ala-Asp-Ala-Lys + Ile-Val-Gly
show the reaction diagram
P98072
21.1% relative activity
-
-
?
Val-Asp-Ala-Asp-Ala-Lys-Ile-Val-Gly + H2O
Val-Asp-Ala-Asp-Ala-Lys + Ile-Val-Gly
show the reaction diagram
P98074
22.5% relative activity
-
-
?
Val-Asp-Asp-Asp-Asp-Lys-2-naphthylamide + H2O
?
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
-
-
?
WDDKG + H2O
?
show the reaction diagram
-
-
-
-
?
WDDRG + H2O
?
show the reaction diagram
-
-
-
-
?
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
p-nitroaniline + Z-Ala-Leu-Arg
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-Phe-Arg-4-methylcoumaryl-7-amide + H2O
7-amino-4-methylcoumarin + Z-Phe-Arg
show the reaction diagram
A4UWM5, -
-
-
-
?
Z-Pro-Phe-Arg-p-nitroanilide + H2O
p-nitroaniline + Z-Pro-Phe-Arg
show the reaction diagram
-
-
-
-
?
MLTAEEKAA + H2O
?
show the reaction diagram
-
Hb 1-9
-
-
?
additional information
?
-
-
the enteropeptidase heavy chain has little influence on the recognition of small peptides, but strongly influences macromolecular substrate recognition
-
-
-
additional information
?
-
-
the catalytic subunit retains the restricted specificity of intact enterokinase but the rate of activation of trypsinogen is much slower
-
-
-
additional information
?
-
-
acidic residues at the P2, P3 and/or P4 position are especially favorable for maximal activity, but are not absolutely necessary
-
-
-
additional information
?
-
-
the highly specific protease cleaves immediately after the carboxyl-terminal residue of the (Asp)4-Lys recognition sequence
-
-
-
additional information
?
-
-
susceptible bonds are either Lys or Arg. The preceding acidic residues could be either Asp, Glu, or carboxymethyl cysteine
-
-
-
additional information
?
-
-
cleaves after Lys residues of peptidyl substrates that resemble trypsinogen activation peptides such as Val-(Asp)4-Lys
-
-
-
additional information
?
-
P98072
Val-(Asp)2-(Ala)2-Lys-Ile-Val-Gly, Val-Ala-Asp-(Ala)2-Lys-Ile-Val-Gly, Val-(Asp)4-Lys-Ile-Val-Gly, Val-(Asp)4-Ala-Ile-Val-Gly and Val-(Asp)4-Glu-Ile-Val-Gly are not hydrolyzed
-
-
-
additional information
?
-
P98074
Val-(Asp)2-(Ala)2-Lys-Ile-Val-Gly, Val-Ala-Asp-(Ala)2-Lys-Ile-Val-Gly, Val-(Asp)4-Lys-Ile-Val-Gly, Val-(Asp)4-Ala-Ile-Val-Gly and Val-(Asp)4-Glu-Ile-Val-Gly are not hydrolyzed
-
-
-
additional information
?
-
-
enterokinase light chain is a serine protease that recognizes Asp-Asp-Asp-Asp-Lys, D4K, sequence and cleaves the C-terminal peptide bond of the lysine residue
-
-
-
additional information
?
-
-
enteropeptidase cleaves the C-terminal end of the substrate recognition sequence Asp-Asp-Asp-Asp-Lys, D4K. Usage of GD4K-conjugated 7-amino-4-methylcoumarin or GD4K-conjugated 2-naphthylamine as a fluorogenic substrates in the assay for enteropeptidase
-
-
-
additional information
?
-
-
high degree of cleavage specificity is exhibited by enteropeptidase
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
Trypsinogen + H2O
?
show the reaction diagram
-
-
-
-
?
Trypsinogen + H2O
?
show the reaction diagram
-
-
-
-
-
Trypsinogen + H2O
?
show the reaction diagram
-
-
-
-
-
Trypsinogen + H2O
?
show the reaction diagram
-
-
-
-
?
Trypsinogen + H2O
?
show the reaction diagram
-
initiation of digestive enzyme activation by converting trypsinogen into trypsin
-
-
-
Trypsinogen + H2O
?
show the reaction diagram
-
initiates activation of pancreatic hydrolases by cleaving and activating trypsinogen
-
-
-
Trypsinogen + H2O
?
show the reaction diagram
-
enterokinase deficiency is a distinct clinical entity characterized by diarrhea, failure to thrive, hypoproteinemia, and edema. Acquired enterokinase deficiency may occur in some diffuse small bowel diseases. Steatorhea of cellac sprue may be due partly to the fact that deficiency of secretin and cholecystokinin may interfere with the action of enterokinase
-
-
-
Trypsinogen + H2O
?
show the reaction diagram
-
the enzyme plays a key role in initiating the proteolytic digestion cascade in duodenum by converting trypsinogen to trypsin
-
-
-
Trypsinogen + H2O
?
show the reaction diagram
-
physiological activator of trypsinogen
-
-
-
Trypsinogen + H2O
?
show the reaction diagram
-
rapid activation of trypsinogen in pancreatic juice. The enzyme has a key permissive role in protein digestion and plays an essential part in the zymogen mechanism that protects the pancreas from self-destruction
-
-
-
trypsinogen + H2O
trypsin
show the reaction diagram
-
-
-
-
?
trypsinogen + H2O
trypsin
show the reaction diagram
P98073
activation by by endoproteolytic cleavage
-
-
?
additional information
?
-
-
enterokinase light chain is a serine protease that recognizes Asp-Asp-Asp-Asp-Lys, D4K, sequence and cleaves the C-terminal peptide bond of the lysine residue
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
50 mM, 3fold activation of hydrolysis of substrates of the general type GD4KX
Ca2+
-
optimal at 0.02mM, inhibitory at 10 mM
Ca2+
-
required
Ca2+
-
required
CoCl2
-
activates
HgCl2
-
activates
K+
-
activating, thiobenzyl benzyloxycarbonyl-L-lysinate as non-specific substrate
Na+
-
activating, thiobenzyl benzyloxycarbonyl-L-lysinate as non-specific substrate
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
alpha-tosyl-lysine chloromethyl ketone
-
-
-
antipain
-
85-95% loss of activtity
antipain
-
-
Aprotinin
-
40-90% loss of activtity
Aprotinin
-
-
Benzamidine
-
75-92% loss of activtity
Benzamidine
-
-
bestatin
-
7% loss of activtity
Ca2+
-
slight inhibition of substrates with one or two Asp/Glu residues at P2-P3 positions
Ca2+
-
50% inhibition at 10 mM
chymostatin
-
20% loss of activtity
diisopropylphosphorofluoridate
-
-
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
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
SDS
-
40-70% loss of activity, thiobenzyl benzyloxycarbonyl-L-lysinate as substrate
Soybean trypsin inhibitor
P98073
-
-
Soybean trypsin inhibitor
-
40-90% loss of activtity
-
trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane
-
-
Triton X-100
-
30-80% loss of activity, thiobenzyl benzyloxycarbonyl-L-lysinate as substrate
Trypsin inhibitor
-
-
-
Trypsin inhibitor
-
basic pancreatic trypsin inhibitor; Kunitz; soybean trypsin inhibitor
-
Trypsin inhibitor
-
Kunitz
-
Trypsin inhibitor
-
bovine pancreatic trypsin inhibitor
-
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
Urea
synthetic construct
-
use of higher amounts of urea (ca. 5 M) has an inhibitory effect on the enzyme activity
Val-(Asp)4-Lys-chloromethane
-
-
Leupeptin
-
94% loss of activtity
additional information
-
EDTA not inhibitory
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Urea
synthetic construct
-
addition of urea (1-4 M) greatly improves enterokinase cleavage specificity at the canonical site and reduces adventitious cleavage
additional information
synthetic construct
-
changing pH, temperature, or the amount of the enzyme has no effect on cleavage pattern
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
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
-
3.3
-
angiotensin II
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
0.0647
-
APFDDDDKIVGG
-
EP light chain
0.353
-
APFDDDDKIVGG
Q6B4R4
at 0.03-0.08 mM, holoenzyme
0.907
-
APFDDDDKIVGG
Q6B4R4
at 0.08-2 mM, holoenzyme
1.06
-
APFDDDDKIVGG
Q6B4R4
at 0.1-3 mM, holoenzyme
1.41
-
APFDDDDKIVGG
Q6B4R4
at 0.03-0.08 mM, EP light chain
0.147
-
APFDDDDRIVGG
Q6B4R4
at 0.014-0.05 mM, holoenzyme
0.3
-
APFDDDGKIVGG
Q6B4R4
at 0.08-2 mM, holoenzyme
1.24
-
benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide
-
-
0.2
-
Boc-Glu(OBzl)-Ala-Arg-MCA
A4UWM5, -
wild-type enzyme, 37C, pH 7.4
1
-
Boc-Glu(OBzl)-Ala-Arg-MCA
A4UWM5, -
E173A mutant, 37C, pH 7.4; P193E mutant, 37C, pH 7.4
1.2
-
Boc-Glu(OBzl)-Ala-Arg-MCA
A4UWM5, -
K63R mutant, 37C, pH 7.4
1.3
-
Boc-Glu(OBzl)-Ala-Arg-MCA
A4UWM5, -
T105E mutant, 37C, pH 7.4
1.2
-
bovine trypsinogen
-
37 C, 25 mM Tris-HCl pH 8.4, 10 mM CaCl2, 0. 4 mM ovomucoid, 0.3 nM enteropeptidase
-
5.6
-
bovine trypsinogen
-
21 C, 50 mM sodium citrate pH 5.6, 1 nM enteropeptidase
-
0.076
-
formyl-Ala-Phe-Arg-4-nitroanilide
-
pH 8.4, 25C
-
0.192
-
formyl-Ala-Phe-Arg-4-nitroanilide
-
pH 8.4, 25C
-
0.167
-
formyl-Ala-Phe-Lys-4-nitroanilide
-
pH 8.4, 25C
-
0.28
-
formyl-Ala-Phe-Lys-4-nitroanilide
-
pH 8.4, 25C
-
0.437
-
G5DKF(NO2)G
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
0.5
0.6
GD4K 2-naphthylamide
-
pH 8.4, 35C
-
0.025
-
GD4K 7-amido-4-methylcoumarin
-
pH 8.4, 35C
-
0.16
-
GD4KF(NO2)G
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
0.16
-
GD4KF(NO2)G
-
50 mM Ca2+, pH 8.0, 37C
0.2
-
GD4KNfa
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
1.12
-
Gly-Asp-ASp-Asp-Asp-2-naphthylamide
-
-
0.034
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
pH 8.4, 37C
0.141
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
pH 8.4, 37C
0.16
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
P98073
pH 8.4, 37C
0.17
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
pH 8.0, 25C
0.2
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
A4UWM5, -
K63R mutant, 37C, pH 7.4
0.22
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
-
0.3
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
A4UWM5, -
E173A mutant, 37C, pH 7.4
0.4
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
A4UWM5, -
P193E mutant, 37C, pH 7.4; T105E mutant, 37C, pH 7.4
0.61
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
-
0.66
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
mutant enzyme K97A
0.7
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
A4UWM5, -
37C, pH 7.4
0.77
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
mutant enzyme K98A
1.25
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
mutant enzyme K96A
0.034
-
Gly-Asp-Asp-Asp-Asp-Lys-naphthylamide
-
in presence of 0.02 mM CaCl2
0.08
-
Gly-Asp-Asp-Asp-Asp-Lys-naphthylamide
-
in presence of 1mM EDTA
0.141
-
Gly-Asp-Asp-Asp-Asp-Lys-naphthylamide
-
in presence of 0.02 mM CaCl2
0.332
-
Gly-Asp-Asp-Asp-Asp-Lys-naphthylamide
-
in presence of 1mM EDTA
0.2
-
glycyl-tetra-L-aspartyl-L-lysine beta-naphthylamide
-
50 mM Ca2+, pH 8.0, 37C
0.525
-
glycyl-tetra-L-aspartyl-L-lysine beta-naphthylamide
-
pH 8.0, 37C
0.0014
-
human cationic trypsinogen
-
37 C, 100 mM Tris-HCl pH 8.0, 1mM CaCl2, 120 nM soybean trypsin inhibitor, 0.13 nM human enteropeptidase
-
0.0072
-
human cationic trypsinogen
-
25 C, 28 mM sodium succinate (pH 5.6), 10 mM CaCl2
-
1.5
-
human cationic trypsinogen
-
37 C, 100 mM Tris-HCl (pH 8.0), 1 mM CaCl2, 120 nM soybean trypsin inhibitor, 0.45 nM enteropeptidase
-
4.2
-
LTAEEKA
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
4
-
MLTAEEKAA
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
596
-
N-alpha-benzyloxycarbonyl-L-lysine thiobenzyl ester hydrochloride
-
37C, pH 8.0
0.076
-
N-formyl-Ala-Phe-Arg-4-nitroanilide
-
-
0.167
-
N-formyl-Ala-Phe-Lys-4-nitroanilide
-
-
0.125
-
PrAD4KP26
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
-
1
-
Pro-Phe-Arg-4-methylcoumaryl-7-amide
A4UWM5, -
E173A mutant, 37C, pH 7.4; P193E mutant, 37C, pH 7.4
1.1
-
Pro-Phe-Arg-4-methylcoumaryl-7-amide
A4UWM5, -
K63R mutant, 37C, pH 7.4
1.3
-
Pro-Phe-Arg-4-methylcoumaryl-7-amide
A4UWM5, -
T105E mutant, 37C, pH 7.4
10
-
Pro-Phe-Arg-4-methylcoumaryl-7-amide
A4UWM5, -
37C, pH 7.4
0.12
-
thiobenzyl benzyloxy-carbonyl-L-lysinate
-
pH 8.4, 25C
0.14
-
thiobenzyl benzyloxy-carbonyl-L-lysinate
P98073
pH 8.4, 37C
0.14
-
thiobenzyl benzyloxy-carbonyl-L-lysinate
-
pH 8.4, 25C
0.05
-
thiobenzyl benzyloxycarbonyl-L-lysinate
-
mutant enzyme R99A
0.1
-
thiobenzyl benzyloxycarbonyl-L-lysinate
-
mutant enzyme K96A
0.12
-
thiobenzyl benzyloxycarbonyl-L-lysinate
-
; mutant enzyme R97A
0.14
-
thiobenzyl benzyloxycarbonyl-L-lysinate
-
-
0.14
-
thiobenzyl benzyloxycarbonyl-L-lysinate
-
mutant enzyme R98A
0.001
-
Trypsinogen
-
bovine guanidinated trypsinogen
0.0045
-
Trypsinogen
-
bovine trypsinogen
0.0056
-
Trypsinogen
-
pH 5.6
0.007
-
Trypsinogen
-
-
0.007
-
Trypsinogen
-
bovine trypsinogen
0.0072
-
Trypsinogen
-
human trypsinogen
0.017
-
Trypsinogen
-
-
0.07
-
Trypsinogen
-
bovine trypsinogen
1.2
-
Trypsinogen
-
pH 8.4
0.72
-
Val-(Ala)2-(Asp)2-Lys-Ile-Val-Gly
P98074
-
8.77
-
Val-(Ala)2-Asp-Ala-Lys-Ile-Val-Gly
P98074
-
1.51
-
Val-(Ala)3-Asp-Lys-Ile-Val-Gly
P98074
-
1.25
-
Val-(Asp)2-Ala-Asp-Lys-Ile-Val-Gly
P98074
-
8.31
-
Val-(Asp)3-Ala-Lys-Ile-Val-Gly
P98074
-
0.8
-
Val-(Asp)3-Glu-Lys-Ile-Val-Gly
P98074
-
0.45
-
Val-(Asp)4-Arg-Ile-Val-Gly
P98074
-
1.77
-
Val-(Asp)4-Lys-Ile-Val-Gly
P98074
-
1.01
-
Val-Ala-(Asp)3-Lys-Ile-Val-Gly
P98074
-
3.39
-
Val-Ala-Asp-Ala-Asp-Lys-Ile-Val-Gly
P98074
-
1.24
-
Val-Asp-(Ala)2-Asp-Lys-Ile-Val-Gly
P98074
-
21.11
-
Val-Asp-(Ala)3-Lys-Ile-Val-Gly
P98074
-
0.76
-
Val-Asp-Ala-(Asp)2-Lys-Ile-Val-Gly
P98074
-
5.57
-
Val-Asp-Ala-Asp-Ala-Lys-Ile-Val-Gly
P98074
-
1.6
-
WDDKG
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
2.1
-
WDDRG
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
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
-
-
0.1
-
Z-Phe-Arg-4-methylcoumaryl-7-amide
A4UWM5, -
37C, pH 7.4; K63R mutant, 37C, pH 7.4; T105E mutant, 37C, pH 7.4
0.2
-
Z-Phe-Arg-4-methylcoumaryl-7-amide
A4UWM5, -
P193E mutant, 37C, pH 7.4
0.4
-
Z-Phe-Arg-4-methylcoumaryl-7-amide
A4UWM5, -
E173A mutant, 37C, pH 7.4
0.265
-
Z-Pro-Phe-Arg-nitroanilide
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
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
-
0.4
-
angiotensin II
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
53
-
benzoyl-Ag ethyl ester
-
-
-
28.4
-
benzoyl-Arg ethyl ester
-
-
262
-
benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide
-
-
4
-
bovine trypsinogen
-
21 C, 50 mM sodium citrate pH 5.6, 1 nM enteropeptidase
-
6.9
-
bovine trypsinogen
-
37 C, 25 mM Tris-HCl pH 8.4, 10 mM CaCl2, 0. 4 mM ovomucoid, 0.3 nM enteropeptidase
-
23.5
-
formyl-Ala-Phe-Arg-4-nitroanilide
-
pH 8.4, 25C
-
26.3
-
formyl-Ala-Phe-Arg-4-nitroanilide
-
pH 8.4, 25C
-
15.4
-
formyl-Ala-Phe-Lys-4-nitroanilide
-
pH 8.4, 25C
-
18.2
-
formyl-Ala-Phe-Lys-4-nitroanilide
-
pH 8.4, 25C
-
0.97
-
G5DKF(NO2)G
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
0.97
-
G5DKF(NO2)G
-
50 mM Ca2+, pH 8.0, 37C
17.33
-
G5DKF(NO2)G
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
17.33
-
G5DKF(NO2)G
-
50 mM Ca2+, pH 8.0, 37C
24.9
-
GD4K 2-naphthylamide
-
pH 8.4, 35C
-
64.8
-
GD4K 7-amido-4-methylcoumarin
-
pH 8.4, 35C
-
0.99
-
GD4KF(NO2)G
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
0.99
-
GD4KF(NO2)G
-
50 mM Ca2+, pH 8.0, 37C
17.83
-
GD4KF(NO2)G
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
17.83
-
GD4KF(NO2)G
-
50 mM Ca2+, pH 8.0, 37C
0.97
-
GD4KNfa
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
16.67
-
GD4KNfa
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
520
-
Gly-Asp-ASp-Asp-Asp-2-naphthylamide
-
-
4.27
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
-
17.1
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
mutant enzyme K96A
20.8
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
pH 8.0, 25C
24.1
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
-
25.5
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
mutant enzyme K97A
39.1
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
mutant enzyme K98A
49.3
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
pH 8.4, 37C
115
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
P98073
pH 8.4, 37C
118
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
pH 8.4, 37C
0.97
-
glycyl-tetra-L-aspartyl-L-lysine beta-naphthylamide
-
50 mM Ca2+, pH 8.0, 37C
16.67
-
glycyl-tetra-L-aspartyl-L-lysine beta-naphthylamide
-
50 mM Ca2+, pH 8.0, 37C
21.5
-
glycyl-tetra-L-aspartyl-L-lysine beta-naphthylamide
-
pH 8.0, 37C
2.3
-
human cationic trypsinogen
-
0.00036 1/sec/mg, incubates 30 min at 37C, pH 9.0, spectrophotometrically measured at 415 nm; 25C, 28 mM sodium succinate (pH 5.6), 10 mM CaCl2
-
8.8
-
human cationic trypsinogen
-
37 C, 100 mM Tris-HCl (pH 8.0), 1 mM CaCl2, 120 nM soybean trypsin inhibitor, 0.45 nM enteropeptidase
-
35.1
-
human cationic trypsinogen
-
37 C, 100 mM Tris-HCl pH 8.0, 1mM CaCl2, 120 nM soybean trypsin inhibitor, 0.13 nM enteropeptidase
-
0.07
-
LTAEEKA
-
pH 8.0, 37C
0.49
-
LTAEEKA
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
25.17
-
MLTAEEKAA
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
111
-
N-alpha-benzyloxycarbonyl-L-lysine thiobenzyl ester
-
37C, pH 8.0
2.62
-
PrAD4KP26
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
-
129
-
thiobenzyl benzyloxy-carbonyl-L-lysinate
-
pH 8.4, 25C
133
-
thiobenzyl benzyloxy-carbonyl-L-lysinate
P98073
pH 8.4, 37C
133
-
thiobenzyl benzyloxy-carbonyl-L-lysinate
-
pH 8.4, 25C
108
-
thiobenzyl benzyloxycarbonyl-L-lysinate
-
mutant enzyme K96A
120
-
thiobenzyl benzyloxycarbonyl-L-lysinate
-
mutant enzyme R99A
128
-
thiobenzyl benzyloxycarbonyl-L-lysinate
-
mutant enzyme R97A; mutant enzyme R98A
129
-
thiobenzyl benzyloxycarbonyl-L-lysinate
-
-
8.1
-
tosyl-Arg methyl ester
-
-
12
-
tosyl-Arg methyl ester
-
-
12
-
Tosyl-Lys methyl ester
-
-
0.005
-
Trypsinogen
-
bovine guanidinated trypsinogen
1.48
-
Trypsinogen
-
bovine trypsinogen
2.3
-
Trypsinogen
-
bovine trypsinogen
2.8
-
Trypsinogen
-
human trypsinogen
4
-
Trypsinogen
-
pH 5.6
4.8
-
Trypsinogen
-
bovine trypsinogen
6.3
-
Trypsinogen
-
-
6.9
-
Trypsinogen
-
pH 8.4
15.9
-
Val-(Ala)2-(Asp)2-Lys-Ile-Val-Gly
P98074
-
14.1
-
Val-(Ala)2-Asp-Ala-Lys-Ile-Val-Gly
P98074
-
17
-
Val-(Ala)3-Asp-Lys-Ile-Val-Gly
P98074
-
14.3
-
Val-(Asp)2-Ala-Asp-Lys-Ile-Val-Gly
P98074
-
14.6
-
Val-(Asp)3-Ala-Lys-Ile-Val-Gly
P98074
-
8
-
Val-(Asp)3-Glu-Lys-Ile-Val-Gly
P98074
-
13.6
-
Val-(Asp)4-Arg-Ile-Val-Gly
P98074
-
20
-
Val-(Asp)4-Lys-Ile-Val-Gly
P98074
-
17.8
-
Val-Ala-(Asp)3-Lys-Ile-Val-Gly
P98074
-
23
-
Val-Ala-Asp-Ala-Asp-Lys-Ile-Val-Gly
P98074
-
28.1
-
Val-Asp-(Ala)2-Asp-Lys-Ile-Val-Gly
P98074
-
9.1
-
Val-Asp-(Ala)3-Lys-Ile-Val-Gly
P98074
-
14.7
-
Val-Asp-Ala-(Asp)2-Lys-Ile-Val-Gly
P98074
-
10.3
-
Val-Asp-Ala-Asp-Ala-Lys-Ile-Val-Gly
P98074
-
0.54
-
WDDKG
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
0.68
-
WDDRG
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
137
-
formyl-Ala-Phe-Arg-4-nitroanilide
-
pH 8.4, 25C
0
309
-
formyl-Ala-Phe-Arg-4-nitroanilide
-
pH 8.4, 25C
0
55
-
formyl-Ala-Phe-Lys-4-nitroanilide
-
pH 8.4, 25C
0
109
-
formyl-Ala-Phe-Lys-4-nitroanilide
-
pH 8.4, 25C
0
42
-
GD4K 2-naphthylamide
-
pH 8.4, 35C
0
2623
-
GD4K 7-amido-4-methylcoumarin
-
pH 8.4, 35C
0
347
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
pH 8.4, 37C
2778
3500
-
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
-
pH 8.4, 37C
2778
950
-
thiobenzyl benzyloxy-carbonyl-L-lysinate
-
pH 8.4, 25C
9730
969
-
thiobenzyl benzyloxy-carbonyl-L-lysinate
-
pH 8.4, 25C
9730
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0000023
-
Soybean trypsin inhibitor
P98073
pH 8.4, 37C
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.045
-
-
-
1.36
-
-
-
220
-
-
thioredoxin-human epidermal growth factor fusion protein as substrate
330
-
-
purified native enzyme, pH 5.0, 25C
2100
-
-
thioredoxin/human epidermal growth factor fusion protein as substrate
8000
-
-
rEKL/His, Gly-Asp-Asp-Asp-Asp-Lys-beta-naphthylamide substrate
9000
-
-
rEKL, Gly-Asp-Asp-Asp-Asp-Lys-beta-naphthylamide substrate
additional information
-
-
-
additional information
-
-
novel assay method for detection of approximately 1 fmol of enzyme. The method depends on quantification of the release of specifically radiolabelled activation peptides from bovine trypsinogen. The assay is applicable to biological fluids such as serum
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
-
-
assay at, substrate trypsin
5.2
5.7
-
bovine trypsinogen
6
9
-
activity rises sharply between pH 5 and 6 and then remains maximal until pH 9
6
-
-
-
6
-
-
immobilised enzyme on paramagnetic microspheres
7.5
-
-
-
8
-
-
free enzyme
8
-
A4UWM5, -
-
8
-
-
assay at
8
-
-
with substrate GD4K 7-amido-4-methylcoumarin
8
-
-
assay at, substrate human interferon-alpha2a
8
-
-
assay at
8
-
-
in presence of 7.5 mM CaCl2
8
-
-
hydrolysis of Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
8.4
-
-
assay at
9
-
-
immobilised enzyme on hexamethylamino Sepabeads
additional information
-
P98074
shows pH optima at 8.0-8.5 for Val-(Asp)4-Lys-Ile-Val-Gly and 8.5-9 for Val-(Asp)3-Ala-Lys-Ile-Val-Gly
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
6.5
-
-
4.5
9.5
-
-
5
10
-
-
6.4
9
-
50% of maximal activity at pH 6.4 and at pH 9.0, hydrolysis of Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
additional information
-
-
-
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
22
-
-
assay at
25
-
-
immobilised enzyme on hexamethylamino Sepabeads or paramagnetic microspheres
25
-
-
assay at
25
-
-
assay at, substrate thiobenzyl benzyloxy-carbonyl-L-lysinate
33
-
-
free enzyme
37
-
-
assay at
37
-
-
assay at, substrate Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
45
-
-
15
40
-
-
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
localized in the brush-border of the enterocytes lining the duodenum and the first 10-20 cm of the small intestine
Manually annotated by BRENDA team
-
duodenal mucosal fluid adhering to the intestinal wall; the enterokinase molecule from the mucosal fluid is identical with that found previously in the mucosal cells but differs from that in the intestinal contents
Manually annotated by BRENDA team
-
duodenal mucosa
Manually annotated by BRENDA team
-
duodenal enterocytes are the major type of cells expressing enterokinase
Manually annotated by BRENDA team
-
duodenal fluid
Manually annotated by BRENDA team
-
mucosa of the first and second portion of the duodenum
Manually annotated by BRENDA team
-
jejunal mucosa
Manually annotated by BRENDA team
-
enterokinase is expressed in the proximal part, the heavy chain anchors the enzyme in the intestinal brush border membrane
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
26000
-
Q6B4R4
SDS-PAGE, MALDI-TOF
36000
-
A4UWM5, -
Western blot
41000
-
-
SDS-PAGE, recombinant enterokinase light chain
43000
-
-
SDS-PAGE, catalytic subunit of Chinese bovine enterokinase
44000
-
-
SDS-PAGE, His-tagged recombinant enterokinase light chain
65000
-
-
SDS-PAGE, represents 28% of total protein
127000
-
-
gradient PAGE
150000
-
-
-
194000
-
-
ultracentrifugation
212000
-
-
gel filtration
296000
-
-
gel filtration
316000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 27000, deglycosylated recombinant enzyme, SDS-PAGE, x * 37000-72000, hyperglycosylated recombinant enzyme, SDS-PAGE
?
-
x * 145000, SDS-PAGE; x * 57000 + x * 82000, SDS-PAGE in presence of 2-mercaptoethanol
?
-
x * 205000-255000, SDS-PAGE
?
-
x * 134000 + x * 62000, SDS-PAGE
?
-
x * 240000 + x * 260000, SDS-PAGE
dimer
-
2 * 62000, SDS-PAGE
heterodimer
-
-
multimer
A4UWM5, -
10 * 44000, gel filtration
trimer
-
-
heterodimer
-
-
additional information
-
the mature active enzyme is comprised of two disulfide-linked polypeptide chains. The heavy chain anchors the enzyme in the intestinal brush border membrane, whereas the light chain represents the catalytic enzyme subunit
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
-
the zymogen proenteropeptidase is activated by trypsin
side-chain modification
-
-
side-chain modification
-
35% carbohydrate
proteolytic modification
-
secreted as a proenzyme, enterokinase converts trypsinogen to trypsin
side-chain modification
-
contains more than 57% carbohydrate per weight
side-chain modification
-
contains 47% neutral sugar and 10% amino sugar
side-chain modification
-
glycoprotein
side-chain modification
-
glycoprotein contains 1.01% hexosamine, 2.17% hexose, 0.36% sialic acid and 0.53% fucose
side-chain modification
-
contains 20% neutral sugar, 20% amino sugars, 15% sialic acids; glycosylated mainly with N-linked carbohydrate chains of the triantennary and tetraantennary complex type
side-chain modification
-
glycosylated mainly with N-linked carbohydrate chains of the triantennary and tetraantennary complex type
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
crystal structure of the enteropeptidase catalytic domain to 2.3 A resolution in complex with the inhibitor Val-(Asp)4-Lys-chloromethane
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
A4UWM5, -
loss of 30% of activity after 4 days
60
-
-
purified recombinant enteropeptidase light chain wild-type and N6D/G21D/G22D/N141D/K209E mutant enteropeptidase light chains show 60% remaining activity
60
-
-
complete destruction of activity towards trypsinogen after 2 min, 60% loss of esterase activity after 8 min. Presence of 4 mM CaCl2 stabilizes both activities
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 ACCESSION NO.
LITERATURE
highly resistant to detergent to triton X-100, tween 20 and SDS
-
no loss of activity at -20C or 4C for 6 months
A4UWM5, -
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
The free enzyme has a half life of 8 days at 23C. The immobilised enzyme has a half life of 29 days at 23C. The immobilised and glutardialdehyde treated enzyme has a half life of 36 days at 23C. The free enzyme has a half life of 85 days at 4C. The immobilised enzyme has a half life of 10 month at 4C.
-
100% activity during storage at 20C for at least for 1.5 years
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
anion exchange chromatography
-
by gel filtration
P98072
His-Tag affinity chromatography
-
Ni2+ affinity chromatography
-
of the recombinant fusion protein by osmotic shock technique and nickel affinity column chromatography
-
of the recombinant fusion protein using nickel affinity column chromatography, refolding and auto-catalytic cleavage
Q6B4R4
recombinant catalytic subunit
-
solubilization of inclusion bodies with 6 M guanidine-HCl, affinity chromatography on STI-agarose
-
KTA-purifier system and a Sepharose QFF column (HiPrep 16/10 QFF)
-
refolded, soluble, and detagged recombinant enteropeptidase light chain by affinity chromatography on soybean trypsin inhibitor agarose
-
solubilization of inclusion bodies with 6 M guanidine-HCl, affinity chromatography on STI-agarose
-
of the recombinant fusion protein using nickel affinity column chromatography
A4UWM5, -
by gel filtration
P98074
native enzyme from duodenal mucosa by anion exchange and benzoamidine affinity chromatography, followed by hydrophobic interaction chromatography and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
catalytic subunit is expressed in escherichia coli BL21
-
expressed in Escherichia coli strain BL21 (DE3)
-
expressed in Pichia pastoris methylotrophic strain GS115
-
expressed in Pichia pastoris strain GS115 mutants Mut+ and MutS
-
expression as fusion protein with thioredoxin in Escherichia coli
-
expression in Escherichia coli as glutathione S-transferase-fusion protein, the gene is a Pro82Arg/Glu176Asp variant of the known bovine EK
Q6B4R4
expression in Escherichia coli BL21
-
expression of a 26300 Da protein containing the catalytic domain of bovine enterokinase, expression in methylotrophic yeast Pichia pastoris
-
expression of catalytic light subunit in Escherichia coli
-
expression of the enterokinase light chain fused to DsbA in Escherichia coli
-
expression of wild-type and mutant enterokinase light chain in Escherichia coli strain Rosetta (DE3)
-
expression as fusion protein with thioredoxin in Escherichia coli
-
expression of light chain of enteropeptidase in Escherichia coli BL21(DE3) in both soluble and insoluble form
P98073
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
-
expression in Escherichia coli
A4UWM5, -
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Y174K
-
site-directed mutagenesis,
Y174K/K99M
-
site-directed mutagenesis, the mutant does not form the active structure
Y174K/K99Q
-
site-directed mutagenesis, the mutantion results in a more site-specific enterokinase light chain
Y174R
-
site-directed mutagenesis,
Y174R/K99M
-
site-directed mutagenesis, the mutant does not form the active structure
Y174R/K99Q
-
site-directed mutagenesis, the mutant does not form the active structure
N6D/G21D/G22D/N141D/K209E
-
site-directed mutagenesis, the mutations lead to supercharging of the protein surface leading to 100fold increased protein solubility
E173A
A4UWM5, -
site directed mutagenesis
E173K
A4UWM5, -
site directed mutagenesis
F144A
A4UWM5, -
site directed mutagenesis
F144S
A4UWM5, -
site directed mutagenesis
K63A
A4UWM5, -
site directed mutagenesis
K63E
A4UWM5, -
site directed mutagenesis
K63R
A4UWM5, -
site directed mutagenesis
P193A
A4UWM5, -
site directed mutagenesis
P193E
A4UWM5, -
site directed mutagenesis
T105A
A4UWM5, -
site directed mutagenesis
T105E
A4UWM5, -
site directed mutagenesis
K99A
-
no cleavage of trypsinogen or Gly-(Asp)4-Lys-beta-naphthylamide and reduced rate of inhibition by Val-(Asp)4-Lys-chloromethane
additional information
-
in order to produce more site-specific enterokinase light chain, EKL, several mutants are generated with substitutions at Tyr174 and Lys99 using the protein disulfide isomerase fusion system. Substitution of Tyr174 by basic residues confers higher specificity on EKL. Mutant enzyme substrate specificity and modeling, overview
additional information
-
improved efficiency of fusion proteins cleavage by enteropeptidase by substitution of the Lys residue by Arg in specific cleavage sequence (Asp)4-Lys
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
additional information
-
even mild protein surface supercharging by mutagenesis can have pronounced effects on protein solubility and stability, overview
additional information
-
improved efficiency of fusion proteins cleavage by enteropeptidase by substitution of the Lys residue by Arg in specific cleavage sequence (Asp)4-Lys
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
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 4C 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 ACCESSION NO.
COMMENTARY
LITERATURE
biotechnology
-
EK is immobilised on hexamethylamino Sepabeads or on amino-modified paramagnetic microspheres. 50% of activity remains after immobilisation
biotechnology
-
purification of 6.8 mg bioactive enzyme from 1l fermentation broth
biotechnology
Q6B4R4
study presents a simple and cost-effective procedure for a large-scale production
synthesis
-
useful tool for in vitro cleavage of fusion proteins
synthesis
-
tool protease in the research and production of gene engineering
synthesis
-
gene engineering studies on processing fusion proteins
synthesis
-
a huge number of therapeutic proteins such as antibodies, coagulation factors, growth hormones or vaccines are produced as fusion proteins. To obtain the therapeutic protein in its monomeric, active form, the fusion partner has to be removed either by chemical or enzymatic cleavage. Enterokinase is a very attractive tool for the in vitro cleavage of fusion proteins
synthesis
-
the enzyme may be useful in amino acid sequence studies for the production of large fragents. The enzyme may also be useful in DNA-recombinant studies in releasing the desired polypeptide chain from neighboring sequences
synthesis
-
the cleavage immediately after the carboxyl-terminal residue of the (Asp)4-Lys recognition sequence allows regeneration of native amino-terminal residues of recombinant proteins, e.g. removal of the thioredoxin and polyhistidine fusion partners from proteins of intrest
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
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
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 specificity of the target site makes enterokinase an ideal tool for cleaving fusion proteins at defined cleavage sites
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
synthesis
P98073
the enzyme can be used for cleavage of fusion proteins due to its high specific activity
analysis
-
enteropeptidase activity is influenced by accessibility of the target site and by downstream sequences
molecular biology
-
the enzyme is used for cleavage of the N-terminal part of recombinant human interferon-alpha2a (IFN-alpha2a) and IFN-alpha2b, expressed in Escherichia coli strains strains BL21 and BL21 (DE3), for production of the protein without the N-terminal methionine residue
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
additional information
-
utility of enterokinase light chain as a site-specific cleavage enzyme is hampered by sporadic cleavage at other sites than the canonical D4K recognition sequence