Information on EC 3.4.21.9 - enteropeptidase

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The expected taxonomic range for this enzyme is: Euteleostomi

EC NUMBER
COMMENTARY hide
3.4.21.9
-
RECOMMENDED NAME
GeneOntology No.
enteropeptidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Activation of trypsinogen by selective cleavage of Lys6-/-Ile bond
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
CAS REGISTRY NUMBER
COMMENTARY hide
9014-74-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
medaka
Swissprot
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
synthetic construct
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
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
APFDDDGKIVGG + H2O
?
show the reaction diagram
N-terminal dodecapeptides of human pancreatitis-associated mutant variant of trypsinogen
-
-
?
APFDDDGRIVGG + H2O
?
show the reaction diagram
N-terminal dodecapeptides of human cationic tryosinogen
-
-
?
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
-
-
?
benzyloxycarbonyl-Phe-Arg-4-methylcoumaryl 7-amide + H2O
?
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
Boc-Glu(OBzl)-Ala-Arg-4-methylcoumaryl-7-amide + H2O
Boc-Glu(OBzl)-Ala-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
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
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
fusion protein Trx/hEGF + H2O
?
show the reaction diagram
fusion protein Trx/hIL-13 + 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
GD4R-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
GDDDDK-2-naphthylamide + H2O
?
show the reaction diagram
-
-
-
-
?
GDDDDK-2-naphthylamide + H2O
GDDDDK + 2-naphthylamine
show the reaction diagram
-
specific substrate
-
-
?
GDDDDK-4-nitroanilide + H2O
GDDDDK + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
glutathione S-transferase-enterokinase + H2O
enterokinase + glutathione S-transferase
show the reaction diagram
GST-EK harboring an EK site
-
-
?
Gly-(L-Asp)4-L-Lys-2-naphthylamide + H2O
?
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
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide + H2O
?
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-2-naphthylamide + H2O
Gly-Asp-Asp-Asp-Asp-Lys + 2-naphthylamine
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
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
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
-
-
?
MLTAEEKAA + H2O
?
show the reaction diagram
-
Hb 1-9
-
-
?
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
-
-
-
?
protein C inhibitor + H2O
?
show the reaction diagram
-
-
-
-
?
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
-
-
?
t-butyloxycarbonyl-E(benzyloxycarbonyl(butanoyl))-Ala-Arg-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
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
3-carboxy-4-nitrophenoxide + ?
show the reaction diagram
thiobenzyl benzyloxy-carbonyl-L-lysinate + H2O
?
show the reaction diagram
thiobenzyl benzyloxycarbonyl-L-lysinate + H2O
3-carboxy-4-nitrothiophenoxide + ?
show the reaction diagram
-
Z-Lys-SBzl
-
-
?
thiobenzyl benzyloxycarbonyl-L-lysinate + H2O
?
show the reaction diagram
-
-
-
-
?
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
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
Val-(Ala)2-(Asp)2-Lys-Ile-Val-Gly + H2O
Val-(Ala)2-(Asp)2-Lys + Ile-Val-Gly
show the reaction diagram
Val-(Ala)2-Asp-Ala-Lys-Ile-Val-Gly + H2O
Val-(Ala)2-Asp-Ala-Lys + Ile-Val-Gly
show the reaction diagram
Val-(Ala)3-Asp-Lys-Ile-Val-Gly + H2O
Val-(Ala)3-Asp-Lys + Ile-Val-Gly
show the reaction diagram
Val-(Asp)2-Ala-Asp-Lys-Ile-Val-Gly + H2O
Val-(Asp)2-Ala-Asp-Lys + Ile-Val-Gly
show the reaction diagram
Val-(Asp)3-Ala-Lys-Ile-Val-Gly + H2O
Val-(Asp)3-Ala-Lys + Ile-Val-Gly
show the reaction diagram
Val-(Asp)3-Glu-Lys-Ile-Val-Gly + H2O
Val-(Asp)3-Glu-Lys + Ile-Val-Gly
show the reaction diagram
Val-(Asp)4-Arg-Ile-Val-Gly + H2O
Val-(Asp)4-Arg + Ile-Val-Gly
show the reaction diagram
Val-(Asp)4-Lys-Ile-Val-Gly + H2O
Val-(Asp)4-Lys + Ile-Val-Gly
show the reaction diagram
Val-Ala-(Asp)2-Ala-Lys-Ile-Val-Gly + H2O
Val-Ala-(Asp)2-Ala-Lys + Ile-Val-Gly
show the reaction diagram
Val-Ala-(Asp)3-Lys-Ile-Val-Gly + H2O
Val-Ala-(Asp)3-Lys + Ile-Val-Gly
show the reaction diagram
Val-Ala-Asp-Ala-Asp-Lys-Ile-Val-Gly + H2O
Val-Ala-Asp-Ala-Asp-Lys + Ile-Val-Gly
show the reaction diagram
Val-Asp-(Ala)2-Asp-Lys-Ile-Val-Gly + H2O
Val-Asp-(Ala)2-Asp-Lys + Ile-Val-Gly
show the reaction diagram
Val-Asp-(Ala)3-Lys-Ile-Val-Gly + H2O
Val-Asp-(Ala)3-Lys + Ile-Val-Gly
show the reaction diagram
Val-Asp-Ala-(Asp)2-Lys-Ile-Val-Gly + H2O
Val-Asp-Ala-(Asp)2-Lys + Ile-Val-Gly
show the reaction diagram
Val-Asp-Ala-Asp-Ala-Lys-Ile-Val-Gly + H2O
Val-Asp-Ala-Asp-Ala-Lys + Ile-Val-Gly
show the reaction diagram
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-Lys-SBzl + H2O
?
show the reaction diagram
-
-
-
-
?
Z-Phe-Arg-4-methylcoumaryl-7-amide + H2O
7-amino-4-methylcoumarin + Z-Phe-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 SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
protein C inhibitor + H2O
?
show the reaction diagram
-
-
-
-
?
Trypsinogen + H2O
?
show the reaction diagram
trypsinogen + H2O
trypsin
show the reaction diagram
trypsinogen + H2O
trypsin + ?
show the reaction diagram
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
COMMENTARY hide
LITERATURE
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
COMMENTARY hide
LITERATURE
IMAGE
alpha-tosyl-lysine chloromethyl ketone
-
-
-
antipain
Aprotinin
basic pancreatic trypsin inhibitor
-
reversible inhibition
-
benzamidine
bestatin
-
7% loss of activtity
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
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
trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane
-
-
Triton X-100
-
30-80% loss of activity, thiobenzyl benzyloxycarbonyl-L-lysinate as substrate
Trypsin inhibitor
Tween-20
-
20-75% loss of activity, thiobenzyl benzyloxycarbonyl-L-lysinate as substrate
Val-(Asp)4-Lys-chloromethane
-
-
additional information
-
EDTA not inhibitory
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
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]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
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 - 1.41
APFDDDDKIVGG
0.147
APFDDDDRIVGG
at 0.014-0.05 mM, holoenzyme
0.3
APFDDDGKIVGG
at 0.08-2 mM, holoenzyme
1.24
benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide
-
-
0.5 - 1.6
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
0.2 - 1.3
Boc-Glu(OBzl)-Ala-Arg-MCA
1.2 - 5.6
bovine trypsinogen
0.076 - 0.192
formyl-Ala-Phe-Arg-4-nitroanilide
0.167 - 0.28
formyl-Ala-Phe-Lys-4-nitroanilide
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
0.2
GD4KNfa
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
0.00258 - 0.018
GD4R-4-nitroanilide
0.019 - 0.6
GDDDDK-2-naphthylamide
0.0179 - 0.276
GDDDDK-4-nitroanilide
0.75
Gly-(L-Asp)4-L-Lys-2-naphthylamide
pH and temperature not specified in the publication
-
1.12
Gly-Asp-ASp-Asp-Asp-2-naphthylamide
-
-
0.034 - 1.25
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
0.08 - 0.332
Gly-Asp-Asp-Asp-Asp-Lys-naphthylamide
0.2 - 0.525
glycyl-tetra-L-aspartyl-L-lysine beta-naphthylamide
0.0014 - 1.5
human cationic trypsinogen
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 - 10
Pro-Phe-Arg-4-methylcoumaryl-7-amide
0.2 - 1.5
t-butyloxycarbonyl-E(benzyloxycarbonyl(butanoyl))-Ala-Arg-7-amido-4-methylcoumarin
0.12 - 0.14
thiobenzyl benzyloxy-carbonyl-L-lysinate
0.05 - 0.14
thiobenzyl benzyloxycarbonyl-L-lysinate
0.001 - 1.2
Trypsinogen
0.72
Val-(Ala)2-(Asp)2-Lys-Ile-Val-Gly
-
8.77
Val-(Ala)2-Asp-Ala-Lys-Ile-Val-Gly
-
1.51
Val-(Ala)3-Asp-Lys-Ile-Val-Gly
-
1.25
Val-(Asp)2-Ala-Asp-Lys-Ile-Val-Gly
-
8.31
Val-(Asp)3-Ala-Lys-Ile-Val-Gly
-
0.8
Val-(Asp)3-Glu-Lys-Ile-Val-Gly
-
0.45
Val-(Asp)4-Arg-Ile-Val-Gly
-
1.77
Val-(Asp)4-Lys-Ile-Val-Gly
-
1.01
Val-Ala-(Asp)3-Lys-Ile-Val-Gly
-
3.39
Val-Ala-Asp-Ala-Asp-Lys-Ile-Val-Gly
-
1.24
Val-Asp-(Ala)2-Asp-Lys-Ile-Val-Gly
-
21.11
Val-Asp-(Ala)3-Lys-Ile-Val-Gly
-
0.76
Val-Asp-Ala-(Asp)2-Lys-Ile-Val-Gly
-
5.57
Val-Asp-Ala-Asp-Ala-Lys-Ile-Val-Gly
-
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
-
-
0.165 - 0.19
Z-Lys-SBzl
0.1 - 0.4
Z-Phe-Arg-4-methylcoumaryl-7-amide
0.265
Z-Pro-Phe-Arg-nitroanilide
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
11.2
A4-mutant trypsinogen
Homo sapiens
-
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
Bos taurus
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
53
benzoyl-Ag ethyl ester
Bos taurus
-
-
-
28.4
benzoyl-Arg ethyl ester
Sus scrofa
-
-
262
benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide
Sus scrofa
-
-
0.36 - 1.81
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
4 - 6.9
bovine trypsinogen
23.5 - 26.3
formyl-Ala-Phe-Arg-4-nitroanilide
15.4 - 18.2
formyl-Ala-Phe-Lys-4-nitroanilide
0.97 - 17.33
G5DKF(NO2)G
24.9
GD4K 2-naphthylamide
Homo sapiens
-
pH 8.4, 35C
64.8
GD4K 7-amido-4-methylcoumarin
Homo sapiens
-
pH 8.4, 35C
0.99 - 17.83
GD4KF(NO2)G
0.97 - 16.67
GD4KNfa
42 - 49
GD4R-4-nitroanilide
17.8 - 115
GDDDDK-2-naphthylamide
122 - 148
GDDDDK-4-nitroanilide
520
Gly-Asp-ASp-Asp-Asp-2-naphthylamide
Sus scrofa
-
-
4.27 - 121
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
0.97 - 21.5
glycyl-tetra-L-aspartyl-L-lysine beta-naphthylamide
2.3 - 35.1
human cationic trypsinogen
0.07 - 0.49
LTAEEKA
25.17
MLTAEEKAA
Bos taurus
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
111
N-alpha-benzyloxycarbonyl-L-lysine thiobenzyl ester
Homo sapiens
-
37C, pH 8.0
2.62
PrAD4KP26
Bos taurus
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
-
0.3 - 1.38
t-butyloxycarbonyl-E(benzyloxycarbonyl(butanoyl))-Ala-Arg-7-amido-4-methylcoumarin
129 - 133
thiobenzyl benzyloxy-carbonyl-L-lysinate
108 - 129
thiobenzyl benzyloxycarbonyl-L-lysinate
8.1 - 12
tosyl-Arg methyl ester
12
Tosyl-Lys methyl ester
Bos taurus
-
-
0.005 - 6.9
Trypsinogen
15.9
Val-(Ala)2-(Asp)2-Lys-Ile-Val-Gly
Sus scrofa
P98074
-
14.1
Val-(Ala)2-Asp-Ala-Lys-Ile-Val-Gly
Sus scrofa
P98074
-
17
Val-(Ala)3-Asp-Lys-Ile-Val-Gly
Sus scrofa
P98074
-
14.3
Val-(Asp)2-Ala-Asp-Lys-Ile-Val-Gly
Sus scrofa
P98074
-
14.6
Val-(Asp)3-Ala-Lys-Ile-Val-Gly
Sus scrofa
P98074
-
8
Val-(Asp)3-Glu-Lys-Ile-Val-Gly
Sus scrofa
P98074
-
13.6
Val-(Asp)4-Arg-Ile-Val-Gly
Sus scrofa
P98074
-
20
Val-(Asp)4-Lys-Ile-Val-Gly
Sus scrofa
P98074
-
17.8
Val-Ala-(Asp)3-Lys-Ile-Val-Gly
Sus scrofa
P98074
-
23
Val-Ala-Asp-Ala-Asp-Lys-Ile-Val-Gly
Sus scrofa
P98074
-
28.1
Val-Asp-(Ala)2-Asp-Lys-Ile-Val-Gly
Sus scrofa
P98074
-
9.1
Val-Asp-(Ala)3-Lys-Ile-Val-Gly
Sus scrofa
P98074
-
14.7
Val-Asp-Ala-(Asp)2-Lys-Ile-Val-Gly
Sus scrofa
P98074
-
10.3
Val-Asp-Ala-Asp-Ala-Lys-Ile-Val-Gly
Sus scrofa
P98074
-
0.54
WDDKG
Bos taurus
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
0.68
WDDRG
Bos taurus
-
in 0.1 M Tris-HCl buffer pH 8.0 at 37C
319 - 354
Z-Lys-SBzl
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.22 - 1.81
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
137 - 309
formyl-Ala-Phe-Arg-4-nitroanilide
55 - 109
formyl-Ala-Phe-Lys-4-nitroanilide
42
GD4K 2-naphthylamide
Homo sapiens
-
pH 8.4, 35C
41937
2623
GD4K 7-amido-4-methylcoumarin
Homo sapiens
-
pH 8.4, 35C
41936
2330 - 18900
GD4R-4-nitroanilide
29.6 - 6052
GDDDDK-2-naphthylamide
537 - 6830
GDDDDK-4-nitroanilide
347 - 3500
Gly-Asp-Asp-Asp-Asp-Lys-2-naphthylamide
0.23 - 2.29
t-butyloxycarbonyl-E(benzyloxycarbonyl(butanoyl))-Ala-Arg-7-amido-4-methylcoumarin
950 - 969
thiobenzyl benzyloxy-carbonyl-L-lysinate
1770 - 2150
Z-Lys-SBzl
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, 37C
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
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
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
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
9
-
immobilised enzyme on hexamethylamino Sepabeads
additional information
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
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 6.5
-
-
4.5 - 9.5
-
-
5 - 10
-
-
6 - 9
-
-
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
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
-
assay at
33
-
free enzyme
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 45
-
-
15 - 40
-
-
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
jejunal mucosa
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26000
SDS-PAGE, MALDI-TOF
36000
-
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
COMMENTARY hide
LITERATURE
heterodimer
multimer
-
10 * 44000, gel filtration
trimer
-
-
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
COMMENTARY hide
LITERATURE
proteolytic modification
side-chain modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure of the enteropeptidase catalytic domain to 2.3 A resolution in complex with the inhibitor Val-(Asp)4-Lys-chloromethane
-
hanging drop vapor diffusion method, using 20% (w/v) polyethylene glycol 10000, 0.1 M HEPES, pH 7.5
-
hanging drop vapor diffusion method, using 20% (w/v) polyethylene glycol 3350, 0.1 M cadmium chloride, 0.1 M sodium acetate (pH 4.6)
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
loss of 30% of activity after 4 days
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
-
no loss of activity at -20C or 4C for 6 months
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
100% activity during storage at 20C for at least for 1.5 years
-
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.
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
anion exchange chromatography
-
by gel filtration
His-Tag affinity chromatography
-
native enzyme from duodenal mucosa by anion exchange and benzoamidine affinity chromatography, followed by hydrophobic interaction chromatography and gel filtration
-
Ni-NTA column 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
-
of the recombinant fusion protein using nickel affinity column chromatography, refolding and auto-catalytic cleavage
recombinant catalytic subunit
-
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
soybean trypsin inhibitor-agarose column chromatography, and gel filtration
-
STI-agarose column chromatography
-
STI-agarose column chromatography and Superdex 200 gel filtration
-
KTA-purifier system and a Sepharose QFF column (HiPrep 16/10 QFF)
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
catalytic subunit is expressed in escherichia coli BL21
-
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli BL21(DE3) codon plus RIL hEPL sc C122S cells
-
expressed in Escherichia coli strain BL21 (DE3)
-
expressed in Escherichia coli strain BL21(DE3) and on M13 filamentous bacteriophage
-
expressed in Pichia pastoris
-
expressed in Pichia pastoris methylotrophic strain GS115
-
expressed in Pichia pastoris strain GS115 mutants Mut+ and MutS
-
expressed in Pichia pastoris X-33 strain
-
expressed in Sf-9 insect cells
expression as fusion protein with thioredoxin in Escherichia coli
expression 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
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 light chain of enteropeptidase in Escherichia coli BL21(DE3) in both soluble and insoluble form
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)
-
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
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K99A
-
no cleavage of trypsinogen or Gly-(Asp)4-Lys-beta-naphthylamide and reduced rate of inhibition by Val-(Asp)4-Lys-chloromethane
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
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
E136Y
-
the mutant shows clearly reduced activity compared to the wild type enzyme
E136Y/R213L
-
the mutant shows about wild type activity
E173A
-
site directed mutagenesis
E173K
-
site directed mutagenesis
F144A
-
site directed mutagenesis
F144S
-
site directed mutagenesis
H24Q
-
the mutant shows slightly reduced activity compared to the wild type enzyme
K63A
-
site directed mutagenesis
K63E
-
site directed mutagenesis
K63R
-
site directed mutagenesis
P193A
-
site directed mutagenesis
P193E
-
site directed mutagenesis
R213L
-
the mutant shows increased activity compared to the wild type enzyme
T105A
-
site directed mutagenesis
T105E
-
site directed mutagenesis
additional information
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
-
fast dilution method in 20 mM NH3 and 10% (v/v) glycerol at pH 10.5 for 72 h at 4?
-
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
COMMENTARY hide
LITERATURE
analysis
biotechnology
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
synthesis
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