Information on EC 3.4.22.8 - clostripain

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The expected taxonomic range for this enzyme is: Clostridium

EC NUMBER
COMMENTARY
3.4.22.8
-
RECOMMENDED NAME
GeneOntology No.
clostripain
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Preferential cleavage: Arg-/-, including Arg-/-Pro, but not Lys-
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
esterification
-
-
-
-
hydrolysis of peptide bond
-
-
endpeptidase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
alpha-clostridipain
-
-
-
-
alpha-clostripain
-
-
alpha-clostripain
Clostridium perfringens JIR325
-
-
-
CLO
Clostridium histolyticum ATCC 19401
P09870
-
-
clostridiopeptidase
-
-
-
-
clostridiopeptidase B
-
-
-
-
Clostridium histolyticum proteinase B
-
-
-
-
clostripain-like protease
-
-
clostripain-like protease
Clostridium perfringens 13
-
-
-
EC 3.4.4.20
-
-
formerly
-
proteinase, Clostridium histolyticum, B
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9028-00-6
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Clostridium histolyticum ATCC 17859
-
-
-
Manually annotated by BRENDA team
Clostridium histolyticum ATCC 17860
-
-
-
Manually annotated by BRENDA team
Clostridium histolyticum ATCC 19401
-
UniProt
Manually annotated by BRENDA team
Clostridium histolyticum ATCC 19401
-
-
-
Manually annotated by BRENDA team
Clostridium histolyticum ATCC 25770
-
-
-
Manually annotated by BRENDA team
Clostridium histolyticum ATCC 6282
-
-
-
Manually annotated by BRENDA team
Clostridium histolyticum ATCC 8034
-
-
-
Manually annotated by BRENDA team
Clostridium perfringens 13
strain 13
-
-
Manually annotated by BRENDA team
Clostridium perfringens JIR325
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-
disruption of the alpha-clostripain gene does not affect alpha-toxin and perfringolysin O production or hemoglobin degradation
malfunction
Clostridium perfringens JIR325
-
disruption of the alpha-clostripain gene does not affect alpha-toxin and perfringolysin O production or hemoglobin degradation
-
physiological function
-
alpha-clostripain is not essential for the pathogenesis of Clostridium perfringens-mediated myonecrosis in mice
physiological function
Clostridium perfringens JIR325
-
alpha-clostripain is not essential for the pathogenesis of Clostridium perfringens-mediated myonecrosis in mice
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
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
Clostridium perfringens, Clostridium perfringens 13
-
-
-
-
?
acyl-L-lysine-4-nitroanilide + H2O
acyl-L-lysine + 4-nitroaniline
show the reaction diagram
Clostridium histolyticum, Clostridium histolyticum ATCC 19401
P09870
-
-
-
?
azocasein + H2O
?
show the reaction diagram
Clostridium perfringens, Clostridium perfringens JIR325
-
-
-
-
?
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
Clostridium perfringens, Clostridium perfringens 13
-
-
-
-
?
benzoyl-Lys-4-nitroanilide + H2O
benzoyl-Lys + 4-nitroaniline
show the reaction diagram
Clostridium perfringens, Clostridium perfringens 13
-
-
-
-
?
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
-
-
-
-
?
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
Clostridium histolyticum, Clostridium histolyticum ATCC 25770, Clostridium histolyticum ATCC 17860, Clostridium histolyticum ATCC 19401, Clostridium histolyticum ATCC 8034, Clostridium histolyticum ATCC 17859, Clostridium histolyticum ATCC 6282
-
-
-
-
?
Nalpha-benzoyl-L-arginine-4-nitroanilide + H2O
Nalpha-benzoyl-L-arginine + 4-nitroaniline
show the reaction diagram
Clostridium histolyticum, Clostridium histolyticum ATCC 19401
P09870
the enzyme exhibits strict specificity for arginine at the P1 position, the enzyme displays 2fold higher specific activity with N-benzoyl-L-arginine-4-nitroanilide compared to acyl-L-lysine-4-nitroanilide
-
-
?
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-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
-
-
-
-
-
p-tosyl-Arg methyl ester + H2O
p-tosyl-Arg + methanol
show the reaction diagram
-
-
-
-
?
p-tosyl-Arg methyl ester + H2O
p-tosyl-Arg + methanol
show the reaction diagram
-
N-alpha-p-tosyl-L-Arg methyl ester
-
-
?
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
-
-
-
-
-
maleyl-Tyr-Arg ethyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
cleaves proteins and synthetic substrates preferentially at the carboxyl group of Arg
-
-
-
additional information
?
-
-
specificity is confined primarily to Arg residues
-
-
-
additional information
?
-
-
the major activity is directed towards the carboxyl peptide linkage of Arg
-
-
-
additional information
?
-
-
clostripain enhanced phagocytosis of apoptotic neutrophils by macrophages
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
enhances esterase activity
Ca2+
-
1 mM, 50% enhancement
Ca2+
-
half-maximal activity at 0.195 mM
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
0.035 mM, 28% inhibition
benzylamidinyl-diazo dichlorotriazine
-
effective protection by N-benzoyl-L-Arg ethyl ester
benzyloxycarbonyl-Phe-Lys-CH2S(CH3)2
-
irreversible
Borate
-
partial inhibition
butylguanidine
-
most effective competitive inhibitor, hydrolysis of benzoyl-Arg ethyl ester
-
butylguanidine
-
-
-
Cd2+
-
0.01 mM, 30% inhibition
citrate
-
partial inhibition
Co2+
-
0.01 mM, 50% inhibition
Cu2+
-
0.01 mM, 30% 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
H2O2
-
complete regeneration by reduction with dithiothreitol
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
-
iodoacetic acid
-
0.035 mM, 13% inhibition
K+
-
100 mM, 40% 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
-
0.035 mM, complete inhibition
Leupeptin
-
potent clostridiopeptidase B-induced kinin release in dog plasma
Leupeptin
-
reversible
macroglobulin
-
alpha2-macroglobulin from human plasma, inhibits both the amidase and protease activity. Macroglobulin from rat and dog plasma
-
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-Lys chloromethyl ketone
-
competitive
N-Alpha-p-tosyl-L-norarginine methyl ester
-
-
N-tosyl-Lys-chloromethylketone
-
0.035 mM, complete inhibition
Na+
-
100 mM, 40% 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
RCM RNase
-
hydrolysis of benzoyl-Arg ethyl ester
-
tosyl-Lys-chloromethane
-
-
Tris
-
partial inhibition
Trypsin inhibitor
-
50% inhibition of amidase activity at 0.04 mM; Kunitz soybean trypsin inhibitor
-
Trypsin inhibitor
-
soy bean trypsin inhibitor, competitive
-
Trypsin inhibitor
-
Kunitz soybean trypsin inhibitor; limabean trypsin inhibitor
-
Veronal
-
partial inhibition
additional information
-
no inhibition by 0.035 PMSF
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
25.4
-
Ac-L-Lys-4-nitroanilide
-
-
0.174
-
alpha-benzoyl-Arg ethyl ester
-
-
0.235
-
alpha-benzoyl-Arg ethyl ester
-
-
0.24
0.25
alpha-benzoyl-Arg ethyl ester
-
depending on assay method
0.26
-
alpha-benzoyl-Arg ethyl ester
-
-
0.16
-
benzoyl-Arg ethyl ester
-
-
0.22
-
benzoyl-Arg ethyl ester
-
-
0.852
-
benzoyl-L-Arg-4-nitroanilide
-
-
1.3
-
N-benzoyl-L-Arg amide
-
-
0.3
-
Nalpha-benzoyl-L-arginine-4-nitroanilide
P09870
purified recombinant enzyme, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25C
0.34
-
Nalpha-benzoyl-L-arginine-4-nitroanilide
P09870
commercial enzyme preparation, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25C
3
-
Nalpha-benzoyl-L-Lys methyl ester
-
-
0.25
-
Nalpha-p-tosyl-L-Arg amide
-
-
0.26
-
p-Nitrophenyl p-guanidinobenzoate
-
-
2.5
-
p-Nitrophenyl p-guanidinobenzoate
-
-
0.022
-
p-toluenesulfonyl-L-Arg methyl ester
-
-
0.8
-
p-toluenesulfonyl-L-Arg methyl ester
-
-
0.25
-
Thr-Ala-Ala
-
-
0.36
-
maleyl-Tyr-Arg ethyl ester
-
-
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]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
22.53
-
Ac-L-Lys-4-nitroanilide
-
-
91
92
alpha-benzoyl-Arg ethyl ester
-
depending on assay method
580
-
alpha-benzoyl-Arg ethyl ester
-
-
643
-
alpha-benzoyl-Arg ethyl ester
-
-
1170
-
benzoyl-Arg ethyl ester
-
-
109.3
-
benzoyl-L-Arg-4-nitroanilide
-
-
87
-
N-benzoyl-L-Arg amide
-
-
38.8
-
Nalpha-benzoyl-L-arginine-4-nitroanilide
P09870
commercial enzyme preparation, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25C
69.5
-
Nalpha-benzoyl-L-arginine-4-nitroanilide
P09870
purified recombinant enzyme, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25C
22
-
Nalpha-benzoyl-L-Lys methyl ester
-
-
10
-
Nalpha-p-tosyl-L-Arg amide
-
-
9.8
-
Nalpha-p-tosyl-L-Arg methyl ester
-
-
31
-
p-Nitrophenyl p-guanidinobenzoate
-
-
86
-
p-toluenesulfonyl-L-Arg methyl ester
-
-
824
-
maleyl-Tyr-Arg ethyl ester
-
-
additional information
-
additional information
-
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]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.887
-
Ac-L-Lys-4-nitroanilide
-
-
302628
128.3
-
benzoyl-L-Arg-4-nitroanilide
-
-
235251
116
-
Nalpha-benzoyl-L-arginine-4-nitroanilide
P09870
commercial enzyme preparation, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25C
284956
234
-
Nalpha-benzoyl-L-arginine-4-nitroanilide
P09870
purified recombinant enzyme, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25C
284956
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.00001
-
histatin 5
-
pH 7.4, 25C
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.15
-
diallyl-2,4,4,9-tetramethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
0.032
-
diallyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
0.09
-
diallyl-7,8-dichloro-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
0.036
-
diallyl-7-benzoyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
0.175
-
dibutyl-2,4,4,9-tetramethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
0.3
-
dibutyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
IC50 above 0.3 mM, in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
0.14
-
dibutyl-7,8-dichloro-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
0.3
-
dibutyl-7-benzoyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo-[b][1,4]diazepin-2-ylphosphonate
-
IC50 above 0.3 mM, in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
0.08
-
diethyl-2,4,4,9-tetramethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
0.49
-
diethyl-2,4,4-trimethyl-8-nitro-2,3,4,5-tetrahydro-1H-benzo-[b][1,4]diazepin-2-ylphosphonate
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
0.22
-
diethyl-7,8-dichloro-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
0.3
-
diethyl-7-benzoyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo-[b][1,4]diazepin-2-ylphosphonate
-
IC50 above 0.3 mM, in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
0.278
-
dimethyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo-[b]-[1,4]diazepin-2-ylphosphonate
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
0.3
-
dimethyl-2,4,4-trimethyl-8-nitro-2,3,4,5-tetrahydro-1H-benzo-[b][1,4]diazepin-2-ylphosphonate
-
IC50 above 0.3 mM, in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
0.78
-
dimethyl-7,8-dichloro-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
0.07
-
dimethyl-7-benzoyl-2,4,4-trimethyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-2-ylphosphonate
-
in 100 mM Tris-HCl buffer, pH 7.4, at 37C
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.022
-
P09870
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 25C
0.039
-
P09870
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 25C
0.119
-
P09870
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 25C
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
-
P09870
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 25C
123.5
-
-
substrate benzoyl-L-Arg-4-nitroanilide
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
hydrolysis of alpha-benzoyl-Arg ethyl ester; in phosphate or Tris-HCl buffer
7.4
7.8
-
hydrolysis of alpha-benzoyl-Arg ethyl ester
7.5
-
-
synthesis of carbobenzoxy-Arg-Pro-NH2
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
38000
-
-
gel filtration
50000
-
-
analytical ultracentrifugation
50000
-
-
equilibrium sedimentation
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 15400, calculation from amino acid analysis
?
-
x * 12500, x * 43000, SDS-PAGE with or without 2-mercaptoethanol
dimer
-
38000, 15000 /16000, SDS-PAGE, 38195, 14916 /15374, TOF mass spectrometry, generated on autocatalytic removal of the propeptide and linker undecapeptide from a single precursor peptide (light chains differ in the N-terminal cleavage site)
dimer
Clostridium perfringens 13
-
38000, 15000 /16000, SDS-PAGE, 38195, 14916 /15374, TOF mass spectrometry, generated on autocatalytic removal of the propeptide and linker undecapeptide from a single precursor peptide (light chains differ in the N-terminal cleavage site)
-
heterodimer
P09870
1 * 38000 + 1 * 15000, SDS-PAGE; 1 * 38061 + 1 * 14929, purified recombinant enzyme, MALDI-TOF mass spectrometry; 1 * 38067 + 1 * 14947, commercial enzyme preparation, MALDI-TOF mass spectrometry; 1 * 38091 + 1 * 14957, purified recombinant enzyme, calculated from amino acid sequence
heterodimer
Clostridium histolyticum ATCC 19401
-
1 * 38000 + 1 * 15000, SDS-PAGE; 1 * 38061 + 1 * 14929, purified recombinant enzyme, MALDI-TOF mass spectrometry; 1 * 38067 + 1 * 14947, commercial enzyme preparation, MALDI-TOF mass spectrometry; 1 * 38091 + 1 * 14957, purified recombinant enzyme, calculated from amino acid sequence
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
Ca2+ depresses thermal inactivation
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Ca2+ stabilizes
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ammonium sulfate precipitation and hydrophobic interaction chromatography
-
ammonium sulfate precipitation, hydroxyapatite column chromatography, and benzamidine-Sepharose column chromatography
P09870
recombinant enzyme
-
cation-exchange chromatography (SP-Sepharose), ultrafiltration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Clostridium perfringens strains 13 and 13DELTA6
P09870
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
-
wild-type and mutant enzyme H176A expressed in Escherichia coli
-
overexpressed in Clostridium perfringens strain 13
-
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
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C231S
-
heterodimer formation is largely impaired
H176A
-
no activity against N-benzoyl-L-Arg ethyl ester
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
efficient biocatalyst for the synthesis of peptide isosteres
synthesis
-
the enzyme can be used for the high yielding synthesis of a variety of peptide bond using L-Arg and, to a lesser degree, L-Lys as substrates with L-amino acid amides or D-amino acid amides
synthesis
-
recombinant clostripain might prove useful in the production of insulin from the proinsulin fusion protein