Information on EC 3.4.22.8 - clostripain

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.22.8
-
RECOMMENDED NAME
GeneOntology No.
clostripain
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Preferential cleavage: Arg-/-, including Arg-/-Pro, but not Lys-
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
esterification
-
-
-
-
hydrolysis of peptide bond
-
-
endpeptidase
-
CAS REGISTRY NUMBER
COMMENTARY hide
9028-00-6
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Ac-L-Lys-4-nitroanilide + H2O
N2-acetyl-L-Lys + 4-nitroaniline
show the reaction diagram
acyl-L-lysine-4-nitroanilide + H2O
acyl-L-lysine + 4-nitroaniline
show the reaction diagram
azocasein + H2O
?
show the reaction diagram
benzoyl-Arg ethyl ester + 3-amino-propionamide
ethanol + benzoyl-Arg-3-amino-propionamide
show the reaction diagram
-
-
-
?
benzoyl-Arg ethyl ester + 3-amino-propionic acid methyl ester
ethanol + benzoyl-Arg-3-amino-propionic acid methyl ester
show the reaction diagram
-
-
-
?
benzoyl-Arg ethyl ester + 4-amino-butyramide
ethanol + benzoyl-Arg-4-amino-butyramide
show the reaction diagram
-
-
-
?
benzoyl-Arg ethyl ester + 4-amino-butyric acid methyl ester
ethanol + benzoyl-Arg-4-amino-butyric acid methyl ester
show the reaction diagram
-
-
-
?
benzoyl-Arg ethyl ester + 5-amino-pentanoic acid
ethanol + benzoyl-Arg-5-amino-pentanoic acid
show the reaction diagram
-
-
-
?
benzoyl-Arg ethyl ester + 5-amino-pentanoic acid amide
ethanol + benzoyl-Arg-5-amino-pentanoic acid amide
show the reaction diagram
-
-
-
?
benzoyl-Arg ethyl ester + 5-amino-pentanoic acid methyl ester
ethanol + benzoyl-Arg-5-amino-pentanoic acid methyl ester
show the reaction diagram
-
-
-
?
benzoyl-Arg ethyl ester + 6-amino-hexanoic acid
ethanol + benzoyl-Arg-6-amino-hexanoic acid
show the reaction diagram
-
-
-
?
benzoyl-Arg ethyl ester + 6-amino-hexanoic acid amide
ethanol + benzoyl-Arg-6-amino-hexanoic acid amide
show the reaction diagram
-
-
-
?
benzoyl-Arg ethyl ester + 6-amino-hexanoic acid methyl ester
ethanol + benzoyl-Arg-6-amino-hexanoic acid methyl ester
show the reaction diagram
-
-
-
?
benzoyl-Arg ethyl ester + 9-amino-nonanoic acid
ethanol + benzoyl-Arg-9-amino-nonanoic acid
show the reaction diagram
-
-
-
?
benzoyl-Arg ethyl ester + 9-amino-nonanoic acid amide
ethanol + benzoyl-Arg-9-amino-nonanoic acid amide
show the reaction diagram
-
-
-
?
benzoyl-Arg ethyl ester + 9-amino-nonanoic acid methyl ester
ethanol + benzoyl-Arg-9-amino-nonanoic acid methyl ester
show the reaction diagram
-
-
-
?
benzoyl-L-Arg-4-nitroanilide + H2O
benzoyl-L-Arg + 4-nitroaniline
show the reaction diagram
benzoyl-Lys-4-nitroanilide + H2O
benzoyl-Lys + 4-nitroaniline
show the reaction diagram
benzoyl-Phe 4-guanidinephenyl ester + 2-amino-4-methyl-pentan-1-ol
4-guanidinephenol + benzoyl-Phe-2-amino-4-methyl-pentan-1-ol
show the reaction diagram
-
-
-
?
benzoyl-Phe 4-guanidinephenyl ester + 3-amino-propane-1,2-diol
4-guanidinephenol + benzoyl-Phe-3-amino-propane-1,2-diol
show the reaction diagram
-
-
-
?
benzoyl-Phe 4-guanidinephenyl ester + 3-amino-propanol
4-guanidinephenol + benzoyl-Phe-3-amino-propanol
show the reaction diagram
-
-
-
?
benzoyl-Phe 4-guanidinephenyl ester + 5-amino-pentanol
4-guanidinephenol + benzoyl-Phe-5-amino-pentanol
show the reaction diagram
-
-
-
?
benzoyl-Phe 4-guanidinephenyl ester + cycloheptylamine
4-guanidinephenol + benzoyl-Phe-cycloheptylamide
show the reaction diagram
-
-
-
?
benzoyl-Phe 4-guanidinephenyl ester + pentylamine
4-guanidinephenol + benzoyl-Phe-pentylamide
show the reaction diagram
-
-
-
?
benzoyl-Phe 4-guanidinephenyl ester + propylamine
4-guanidinephenol + benzoyl-Phe-propylamide
show the reaction diagram
-
-
-
?
carbobenzoxy-Arg methyl ester + L-Pro-NH2
carbobenzoxy-Arg-Pro-NH2 + methanol
show the reaction diagram
-
no activity with carbobenzoxy-D-Arg methyl ester
-
?
carbobenzoxy-L-Arg methyl ester + D-Phe-NH2
carbobenzoxy-Arg-D-Phe-NH2 + methanol
show the reaction diagram
-
no activity with carbobenzoxy-D-Arg methyl ester
-
-
?
carbobenzoxy-L-Arg methyl ester + D-Trp-NH2
carbobenzoxy-Arg-D-Trp-NH2 + methanol
show the reaction diagram
-
no activity with carbobenzoxy-D-Arg methyl ester
-
-
?
carbobenzoxy-L-Arg methyl ester + Gly-NH2
carbobenzoxy-Arg-Gly-NH2 + methanol
show the reaction diagram
-
no activity with carbobenzoxy-D-Arg methyl ester
-
-
?
carbobenzoxy-L-Arg methyl ester + L-Phe-NH2
carbobenzoxy-Arg-Phe-NH2 + methanol
show the reaction diagram
-
no activity with carbobenzoxy-D-Arg methyl ester
-
-
?
carbobenzoxy-L-Arg methyl ester + L-Trp-NH2
carbobenzoxy-Arg-Trp-NH2 + methanol
show the reaction diagram
-
no activity with carbobenzoxy-D-Arg methyl ester
-
-
?
cholecystokinin + H2O
cholecystokinin 8
show the reaction diagram
-
cleavage of large forms of cholecystokinin
-
?
histone H3 + H2O
?
show the reaction diagram
-
-
-
-
?
histone H4 + H2O
?
show the reaction diagram
-
-
-
-
?
L-Arg methyl ester + H2O
L-Arg + methanol
show the reaction diagram
-
-
-
-
?
maleyl-Tyr-Arg ethyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzoyl-Arg amide + H2O
N-benzoyl-Arg + NH3
show the reaction diagram
-
-
-
-
?
N-benzoyl-Arg benzyl ester + H2O
N-benzoyl-Arg + benzyl alcohol
show the reaction diagram
-
-
-
-
?
N-benzoyl-Arg ethyl ester + H2O
N-benzoyl-Arg + ethanol
show the reaction diagram
-
-
-
-
?
N-benzoyl-Arg isopropyl ester + H2O
N-benzoyl-Arg isopropanol
show the reaction diagram
-
-
-
-
?
N-benzoyl-Arg methyl ester + H2O
N-benzoyl-Arg + methanol
show the reaction diagram
-
-
-
-
?
N-benzoyl-Arg naphthylamide + H2O
N-benzoyl-Arg + naphthylamine
show the reaction diagram
-
-
-
-
?
N-benzoyl-DL-arginine 4-nitroanilide + H2O
N-benzoyl-DL-arginine + 4-nitroaniline
show the reaction diagram
-
-
-
?
N-Benzoyl-L-Arg ethyl ester + H2O
N-Benzoyl-L-Arg + ethanol
show the reaction diagram
-
-
-
?
Nalpha-benzoyl-Arg p-nitroanilide + H2O
Nalpha-benzoyl-Arg + p-nitroaniline
show the reaction diagram
-
-
-
-
?
Nalpha-benzoyl-DL-arginine-4-nitroanilide + H2O
Nalpha-benzoyl-DL-arginine + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Nalpha-benzoyl-L-arginine ethyl ester + H2O
?
show the reaction diagram
Nalpha-benzoyl-L-arginine-4-nitroanilide + H2O
Nalpha-benzoyl-L-arginine + 4-nitroaniline
show the reaction diagram
Nalpha-benzoyl-Lys methyl ester + H2O
Nalpha-benzoyl-Lys + methanol
show the reaction diagram
-
-
-
-
?
Nalpha-p-tosyl-Arg amide + H2O
Nalpha-p-tosyl-Arg + NH3
show the reaction diagram
-
-
-
-
?
p-nitrophenyl p-guanidinobenzoate + H2O
p-nitrophenol + p-guanidinobenzoate
show the reaction diagram
p-tosyl-Arg methyl ester + H2O
p-tosyl-Arg + methanol
show the reaction diagram
parvalbumin + H2O
?
show the reaction diagram
-
exclusive cleavage of the Arg75-Ala76 bond
-
-
?
proinsulin fusion protein + H2O
insulin + ?
show the reaction diagram
-
clostripain specifically cleaves the arginine residues in the leader peptide and in the C-peptide
-
-
?
succinyl-CoA synthetase + H2O
?
show the reaction diagram
-
the mutant forms of E. coli succinyl-CoA synthetase, W76F and W43,76,248F are more sensitive to proteolysis by clostripain than the wild-type enzyme. No influence of phosphorylation or dephosphorylation state. Argbeta80 is the principal site of inactivation by clostripain
-
-
?
Thr-Ala-Ala + H2O
?
show the reaction diagram
-
-
-
-
-
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5,5'-dithiobis(2-nitrobenzoate)
-
-
ADP
-
proteolysis of succinyl-CoA synthetase
antipain
-
potent clostridiopeptidase B-induced kinin release in dog plasma
ATP
-
proteolysis of succinyl-CoA synthetase
benzamidine
benzylamidinyl-diazo dichlorotriazine
-
effective protection by N-benzoyl-L-Arg ethyl ester
benzyloxycarbonyl-Phe-Lys-CH2S(CH3)2
-
irreversible
Borate
-
partial inhibition
Butylamine
-
-
butylguanidine
-
citrate
-
partial inhibition
diallyl-2,4,4,9-tetramethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
-
diallyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
-
diallyl-7,8-dichloro-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
-
diallyl-7-benzoyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
-
dibutyl-2,4,4,9-tetramethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
-
dibutyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
-
dibutyl-7,8-dichloro-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
-
dibutyl-7-benzoyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo-[b][1,4]diazepin-2-ylphosphonate
-
-
diethyl-2,4,4,9-tetramethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
-
diethyl-2,4,4-trimethyl-8-nitro-2,3,4,5-tetrahydro-1H-benzo-[b][1,4]diazepin-2-ylphosphonate
-
-
diethyl-7,8-dichloro-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
-
diethyl-7-benzoyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo-[b][1,4]diazepin-2-ylphosphonate
-
-
diethyldicarbonate
-
-
diisopropylfluorophosphate
-
-
dimethyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo-[b]-[1,4]diazepin-2-ylphosphonate
-
-
dimethyl-2,4,4-trimethyl-8-nitro-2,3,4,5-tetrahydro-1H-benzo-[b][1,4]diazepin-2-ylphosphonate
-
-
dimethyl-7,8-dichloro-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
-
dimethyl-7-benzoyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
-
E-64
-
0.035 mM, 68% inhibition
ethylamine
-
-
Ethylguanidine
-
-
histatin 5
-
23.6 nM, 50% inhibition, competitive. The salivary protein could be used for the prevention or treatment of topical infactions such as those caused by Clostridium histolyticum
-
iodoacetamide
-
-
iodoacetic acid
-
0.035 mM, 13% inhibition
L-1-chloro-3-[4-tosylamido]-7-amino-2-heptanone
-
complete inhibition
L-3-carboxy-2,3-trans-epoxypropionyl-leucylamido-(4-guanidino)butane
-
i.e. compounds E-64
leupeptin
macroglobulin
-
alpha2-macroglobulin from human plasma, inhibits both the amidase and protease activity. Macroglobulin from rat and dog plasma
-
methylamine
-
-
Methylguanidine
-
-
N-alpha-p-nitrobenzyloxycarbonyl-Arg chloromethyl ketone
-
irreversible
N-Alpha-p-tosyl-L-homoarginine methyl ester
-
-
N-alpha-p-tosyl-L-Lys chloromethyl ketone
N-Alpha-p-tosyl-L-norarginine methyl ester
-
-
N-tosyl-Lys-chloromethylketone
-
0.035 mM, complete inhibition
Nalpha-tosyl-L-lysine chloromethylketone
-
complete inhibition at 0.01 mg/ml
ovoinhibitor
-
0.05 mM, 55% reduced amidase activity
-
phenylmethylsulfonyl fluoride
-
complete inhibition
Polyarginine
-
hydrolysis of benzoyl-Arg ethyl ester
polylysine
-
hydrolysis of benzoyl-Arg ethyl ester
Propylamine
-
-
propylguanidine
-
-
RCM RNase
-
hydrolysis of benzoyl-Arg ethyl ester
-
tosyl-Lys-chloromethane
-
-
Tris
-
partial inhibition
Trypsin inhibitor
-
Veronal
-
partial inhibition
additional information
-
no inhibition by 0.035 PMSF
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
25.4
Ac-L-Lys-4-nitroanilide
-
-
0.174 - 0.26
alpha-benzoyl-Arg ethyl ester
0.16 - 0.22
benzoyl-Arg ethyl ester
0.852
benzoyl-L-Arg-4-nitroanilide
-
-
0.36
maleyl-Tyr-Arg ethyl ester
-
-
1.3
N-benzoyl-L-Arg amide
-
-
0.3 - 0.34
Nalpha-benzoyl-L-arginine-4-nitroanilide
3
Nalpha-benzoyl-L-Lys methyl ester
-
-
0.25
Nalpha-p-tosyl-L-Arg amide
-
-
0.26 - 2.5
p-Nitrophenyl p-guanidinobenzoate
0.022 - 0.8
p-toluenesulfonyl-L-Arg methyl ester
0.25
Thr-Ala-Ala
-
-
additional information
additional information
-
effect of pH-value on Km-value for the hydrolysis of alpha-benzoyl-Arg ethyl ester, effect of temperature on Km-values
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
22.53
Ac-L-Lys-4-nitroanilide
Clostridium perfringens
-
-
91 - 643
alpha-benzoyl-Arg ethyl ester
1170
benzoyl-Arg ethyl ester
Hathewaya histolytica
-
-
109.3
benzoyl-L-Arg-4-nitroanilide
Clostridium perfringens
-
-
824
maleyl-Tyr-Arg ethyl ester
Hathewaya histolytica
-
-
87
N-benzoyl-L-Arg amide
Hathewaya histolytica
-
-
38.8 - 69.5
Nalpha-benzoyl-L-arginine-4-nitroanilide
22
Nalpha-benzoyl-L-Lys methyl ester
Hathewaya histolytica
-
-
10
Nalpha-p-tosyl-L-Arg amide
Hathewaya histolytica
-
-
9.8
Nalpha-p-tosyl-L-Arg methyl ester
Hathewaya histolytica
-
-
31
p-Nitrophenyl p-guanidinobenzoate
Hathewaya histolytica
-
-
86
p-toluenesulfonyl-L-Arg methyl ester
Hathewaya histolytica
-
-
additional information
additional information
Hathewaya histolytica
-
effect of pH-value on the turnover-number for hydrolysis of alpha-benzoyl-Arg ethyl ester, effect of temperature on turnover numbers
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.887
Ac-L-Lys-4-nitroanilide
Clostridium perfringens
-
-
40630
128.3
benzoyl-L-Arg-4-nitroanilide
Clostridium perfringens
-
-
24211
116 - 234
Nalpha-benzoyl-L-arginine-4-nitroanilide
14730
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00001
histatin 5
-
pH 7.4, 25°C
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.15
diallyl-2,4,4,9-tetramethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
0.032
diallyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
0.09
diallyl-7,8-dichloro-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
0.036
diallyl-7-benzoyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
0.175
dibutyl-2,4,4,9-tetramethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
0.3
dibutyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
IC50 above 0.3 mM, in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
0.14
dibutyl-7,8-dichloro-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
0.3
dibutyl-7-benzoyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo-[b][1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
IC50 above 0.3 mM, in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
0.08
diethyl-2,4,4,9-tetramethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
0.49
diethyl-2,4,4-trimethyl-8-nitro-2,3,4,5-tetrahydro-1H-benzo-[b][1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
0.22
diethyl-7,8-dichloro-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
0.3
diethyl-7-benzoyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo-[b][1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
IC50 above 0.3 mM, in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
0.278
dimethyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo-[b]-[1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
0.3
dimethyl-2,4,4-trimethyl-8-nitro-2,3,4,5-tetrahydro-1H-benzo-[b][1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
IC50 above 0.3 mM, in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
0.78
dimethyl-7,8-dichloro-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
0.07
dimethyl-7-benzoyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
Hathewaya histolytica
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.022
commercial enzyme preparation, using acyl-L-lysine-4-nitroanilide as substrate, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25°C
0.039
purified recombinant enzyme, using acyl-L-lysine-4-nitroanilide as substrate, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25°C
0.119
culture supernatant, using Nalpha-benzoyl-L-arginine-4-nitroanilide as substrate, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25°C
0.172
-
substrate benzoyl-L-Lys-4-nitroanilide
4.28
-
substrate benzoyl-L-Arg-4-nitroanilide
25.45
-
substrate Ac-L-Lys-4-nitroanilide
61.2
recombinant enzyme after 515fold purification, using Nalpha-benzoyl-L-arginine-4-nitroanilide as substrate, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25°C
123.5
-
substrate benzoyl-L-Arg-4-nitroanilide
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5
-
substrates: Bz-L-Arg-p-nitroanilide and Ac-L-Lys-4-nitroanilide
7 - 7.2
-
hydrolysis of alpha-N-benzoyl-Arg ethyl ester
7
-
hydrolysis of alpha-benzoyl-Arg ethyl ester
7.2
-
hydrolysis of L-Arg methyl ester, in phosphate buffer
7.3
-
synthesis of carbobenzoxy-Arg-Gly-NH2
7.4 - 7.8
7.5
-
synthesis of carbobenzoxy-Arg-Pro-NH2
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8
-
pH 6.0: about 35% of maximal activity, pH 8.5: about 70% of maximal activity, synthesis of carbobenzoxy-Arg-Gly-NH2
6.5 - 8.5
-
about 35% of maximal activity at pH 6.5 and at pH 8.5, hydrolysis of benzoyl-Arg ethyl ester
7 - 8.5
-
pH 7.0: about 40% of maximal activity, pH 8.5: about 50% of maximal activity, synthesis of carbobenzoxy-Arg-Pro-NH2
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
12500
-
x * 12500, x * 43000, SDS-PAGE with or without 2-mercaptoethanol
14929
1 * 38061 + 1 * 14929, purified recombinant enzyme, MALDI-TOF mass spectrometry
14947
1 * 38067 + 1 * 14947, commercial enzyme preparation, MALDI-TOF mass spectrometry
14957
1 * 38091 + 1 * 14957, purified recombinant enzyme, calculated from amino acid sequence
15000
1 * 38000 + 1 * 15000, SDS-PAGE
15400
-
x * 15400, calculation from amino acid analysis
38061
1 * 38061 + 1 * 14929, purified recombinant enzyme, MALDI-TOF mass spectrometry
38067
1 * 38067 + 1 * 14947, commercial enzyme preparation, MALDI-TOF mass spectrometry
38091
1 * 38091 + 1 * 14957, purified recombinant enzyme, calculated from amino acid sequence
43000
-
x * 12500, x * 43000, SDS-PAGE with or without 2-mercaptoethanol
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterodimer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
-
the clostripain core protein is composed of the light and the heavy chain subunits linked by a nonapeptide into a single polypeptide chain. Linker removal is due to autocatalytic processing yielding active heterodimeric enzyme. The linker nonapeptide serves as an important transient intramolecular inhibitor in the cellular self-defense program evolved by the natural host Clostridium histolyticum
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
Ca2+ depresses thermal inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Ca2+ stabilizes
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation and hydrophobic interaction chromatography
-
ammonium sulfate precipitation, hydroxyapatite column chromatography, and benzamidine-Sepharose column chromatography
cation-exchange chromatography (SP-Sepharose), ultrafiltration
-
recombinant enzyme
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
clostripain gene is modified and its signal sequence is replaced with that of penicillin G acylase. The core clostripain protein fused to the penicillin G acylase signal peptide is also prepared. With regard to the expression of the clostripain precursors, the majority of clostripain activity is observed in the culture media, thereby indicating that both the clostripain signal peptide and the penicillin G acylase signal peptide are recognized in the Escherichia coli secretion pathway, and the precursors successfully mature into the active form. The activity is rather low when the core protein is expressed, which indicates that the clostripain pro-peptide is important in the formation of the active enzyme in Escherichia coli
-
expressed in Clostridium perfringens strains 13 and 13DELTA6
expression in Bacillus subtilis, the Clostridium histolyticum signal peptide is efficiently recognized by the Bacillus subtilis secretion apparatus; expression in Escherichia coli; the enzyme is not transported to the Escherichia coli periplasm
-
expression in Escherichia coli
-
overexpressed in Clostridium perfringens strain 13
-
wild-type and mutant enzyme H176A expressed in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C231S
-
heterodimer formation is largely impaired
H176A
-
no activity against N-benzoyl-L-Arg ethyl ester
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
-
influence of protease inhibitors aprotinin, phenylmethylsulfonyl fluoride, L-1-chloro-3-[4-tosylamido]-7-amino-2-heptanone, and chymostatin on toxicity of Clostridium histolyticum supernatant towards HeLa cells. Aprotinin has no effect, while the other inhibitors potentiate the cytotoxic activity of Clostridium histolyticum probybly by hindering the natural defense of cells. In addition, phenylmethylsulfonyl fluoride and L-1-chloro-3-[4-tosylamido]-7-amino-2-heptanone block clostripain enzymic activity
synthesis