Information on EC 3.4.22.B49 - cathepsin L1

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.22.B49
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
cathepsin L1
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
clear preference for Arg at P1 position (Lys, Glu, Thr, and Met are less efficient). FheCL1 shows distinct preference for hydrophobic amino acids in the P2, Leu is favored. Cathepsin L1 can accommodate Pro in the P2 position, but less efficiently than cathepsin L2. FheCL1 produces clear degradation fragments from collagen
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
60616-82-2
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
olive flounder
-
-
Manually annotated by BRENDA team
pearl oyster
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
benzoyl-Phe-Val-Arg-4-methylcoumarinyl-7-amide + H2O
benzoyl-Phe-Val-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
cathepsin L1
-
-
?
benzoyl-Phe-Val-Arg-7-amido-4-methylcoumarin + H2O
benzoyl-Phe-Val-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Arg-4-methylcoumarinyl-7-amide + H2O
benzyloxycarbonyl-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
cathepsin L1
-
-
?
benzyloxycarbonyl-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Arg-Arg-4-methylcoumarinyl-7-amide + H2O
benzyloxycarbonyl-Arg-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
cathepsin L1
-
-
?
benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Arg-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-L-Leu-L-Arg-4-methylcoumarinyl-7-amide + H2O
benzyloxycarbonyl-L-Leu-L-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
FheCL1
-
-
?
benzyloxycarbonyl-L-Phe-L-Arg-4-methylcoumarinyl-7-amide + H2O
benzyloxycarbonyl-L-Phe-L-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
benzyloxycarbonyl-L-phenylalanyl-L-arginine 4-methylcoumarinyl-7-amide + H2O
?
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-L-Pro-L-Arg-4-methylcoumarinyl-7-amide + H2O
benzyloxycarbonyl-L-Pro-L-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
FheCL1
-
-
?
benzyloxycarbonyl-Leu-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Leu-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Phe-Arg-4-methylcoumarinyl-7-amide + H2O
benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
cleaved by cathepsin L2 with much greater affinity than by cathepsin L1
-
-
?
benzyloxycarbonyl-Phe-Arg-4-nitroanilide + H2O
benzyloxycarbonyl-Phe-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
benzyloxycarbonyl-Phe-Arg-7-amido-4-trifluoromethylcoumarin + H2O
benzyloxycarbonyl-Phe-Arg + 7-amino-4-trifluoromethylcoumarin
show the reaction diagram
-
best substrate
-
-
?
benzyloxycarbonyl-Phe-Phe-Arg-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
renatured CPFhW
-
-
?
benzyloxycarbonyl-Pro-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Pro-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
Collagen + H2O
?
show the reaction diagram
whereas FheCL1 produces clear degradation fragments, FheCL2 degrades the collagen completely, particularly at pH 4.0, indicating that only the latter cleaves efficiently within the helical structures
-
-
?
Collagen type I + H2O
?
show the reaction diagram
-
-
-
?
Collagen type III + H2O
?
show the reaction diagram
-
-
-
?
collagen type IV + H2O
?
show the reaction diagram
-
-
-
?
Fibronectin + H2O
?
show the reaction diagram
-
-
-
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
-
?
H-Leu-Val-Tyr-4-methylcoumarinyl-7-amide + H2O
H-Leu-Val-Tyr + 7-amino-4-methylcoumarin
show the reaction diagram
-
cleaved by cathepsin L2 with much greater affinity than by cathepsin L1
-
-
?
Hemoglobin + H2O
?
show the reaction diagram
human IgG + H2O
?
show the reaction diagram
-
both cathepsins L produce similar degradation patterns and cleave all human IgG subclasses at the hinge region, yielding at pH 7.3 and 37°C Fab and Fc fragments in the case of IgG1 and IgG3 or Fab(2) and Fc in IgG2 and IgG4. Both liver fluke cathepsins L cleave the peptide bonds 237His-Thr, 237Glu-Cys, 233Gly-Asp, and 241Ser-Cys of the gamma1, gamma2, gamma3, and gamma4 H chains, respectively. Therefore, the enzymes are interacting with the following P3-P'3 sequences, Lys-Thr-His-Thr-Cys-Pro, Cys-Val-Glu-Asp-Pro-Pro, Pro-Leu-Gly-Asp-Thr-Thr, and Cys-Pro-Ser-Cys-Pro-Ala. The specificity of the liver fluke cathepsins L for peptide bonds in proteins is less defined. The P1 position, for instance, can be occupied by hydrophobic, hydrophilic, acidic, or basic residues. The P3 and P2 positions are occupied by hydrophobic amino acids with the exception of the gamma1 sequence which contains a basic lysine and a hydrophilic threonine, respectively. In addition the specificity between the enzyme and its substrate would depend on which of the amino acids of the substrate can be really exposed to the active site
-
-
?
Leu-Val-Tyr-7-amido-4-methylcoumarin + H2O
Leu-Val-Tyr + 7-amino-4-methylcoumarin
show the reaction diagram
-
cleaved by cathepsin L2 with much greater affinity than by cathepsin L1
-
-
?
ovalbumin + H2O
?
show the reaction diagram
procathepsin L1 + H2O
?
show the reaction diagram
-
procathepsin L1 autocatalytically processes and activates to its mature enzyme (FheCL1) over a wide pH range 4.0-7.3. Activation is more rapid at low pH. Maturation initiates with cleavages of a small proportion of molecules within the central region of the prosegment, possibly by intramolecular events. Activation to fully mature enzymes is achieved by a precise intermolecular cleavage at a Leu12-Ser11-/-His10 sequence within the nonconserved C-terminal region of the prosegment. Active site variant FheproCL1C26G and a double variant FheproCL1L12P/C26G cannot autocatalytically process. The former is susceptible to trans-processing at a Leu12-Ser11-/-His10 sequence by preactivated FheCL1, but the latter is not
-
-
?
succinyl-Ala-Phe-Lys-4-methylcoumarinyl-7-amide + H2O
succinyl-Ala-Phe-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
cathepsin L1
-
-
?
succinyl-Ala-Phe-Lys-7-amido-4-methylcoumarin + H2O
tosyl-Ala-Phe-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
succinyl-Leu-Leu-Val-Tyr-4-methylcoumarinyl-7-amide + H2O
succinyl-Leu-Leu-Val-Tyr + 7-amino-4-methylcoumarin
show the reaction diagram
-
cathepsin L1
-
-
?
succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin + H2O
succinyl-Leu-Leu-Val-Tyr + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
t-butyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin + H2O
t-butyloxycarbonyl-Val-Leu-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
t-butyloxycarbonyl-Val-Pro-Arg-7-amido-4-methylcoumarin + H2O
t-butyloxycarbonyl-Val-Pro-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
cleaved by cathepsin L2 with much greater affinity than by cathepsin L1
-
-
?
tert-butoxycarbonyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide + H2O
tert-butoxycarbonyl-Gly-Pro-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
FheCL1
-
-
?
tert-butyloxycarbonyl-Val-Leu-Lys-4-methylcoumarinyl-7-amide + H2O
tert-butyloxycarbonyl-Val-Leu-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
cathepsin L1
-
-
?
tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumarinyl-7-amide + H2O
tert-butyloxycarbonyl-Val-Pro-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
cleaved by cathepsin L2 with much greater affinity than by cathepsin L1
-
-
?
tosyl-Ala-Phe-Lys-4-methylcoumarinyl-7-amide + H2O
tosyl-Ala-Phe-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
cathepsin L1
-
-
?
tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide + H2O
tosyl-Gly-Pro-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin + H2O
tosyl-Gly-Pro-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
cleaved by cathepsin L2 with much greater affinity than by cathepsin L1
-
-
?
tosyl-Gly-Pro-Lys-4-methylcoumarinyl-7-amide + H2O
tosyl-Gly-Pro-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
cleaved by cathepsin L2 with much greater affinity than by cathepsin L1
-
-
?
tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin + H2O
tosyl-Gly-Pro-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
cleaved by cathepsin L2 with much greater affinity than by cathepsin L1
-
-
?
Z-Phe-Arg-OMe + SerNH2
Z-Phe-Arg-Ser-NH2 + methanol
show the reaction diagram
-
peptide synthesis
-
-
?
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
Collagen type I + H2O
?
show the reaction diagram
Q9GRW5
-
-
-
?
Collagen type III + H2O
?
show the reaction diagram
Q9GRW5
-
-
-
?
collagen type IV + H2O
?
show the reaction diagram
Q9GRW5
-
-
-
?
Fibronectin + H2O
?
show the reaction diagram
Q9GRW5
-
-
-
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
-
?
Hemoglobin + H2O
?
show the reaction diagram
additional information
?
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Aprotinin
-
complete inhibition at 50 nM at a molar ratio of enzyme to inhibitor of 17:1; inhibits enzyme activity almost completely at a molar ratio of enzyme to inhibitor of 17:1
benzyloxycarbonyl-L-phenylalanyl-L-alanine-diazomethylketone(-CHN2)
-
-
Benzyloxycarbonyl-Phe-Ala-CHN2
-
-
benzyloxycarbonyl-Phe-Ala-diazomethyl ketone
-
FheCL1
CA074
-
25.3% inhibition at 0.05 mM
cathepsin K inhibitor II
-
FheCL1
-
chymostatin
-
85.6% inhibition at 0.05 mM
Cystatin
-
47.5% inhibition at 0.05 mM
-
E-64
-
i.e. L-trans-epoxysuccinyl-leucyamido(4-guanidino) butane, 26.6% inhibition at 0.05 mM
N-2,3,4,5,6-pentafluorobenzoyl-L-leucyl-glycine nitrile
-
-
N-2,3,4,5-tetrafluorobenzoyl-L-leucyl-glycine nitrile
-
-
N-2,3,4-trifluorobenzoyl-L-leucyl-glycine nitrile
-
-
N-2,3,6-trifluorobenzoyl-L-leucyl-glycine nitrile
-
-
N-2,3-difluorobenzoyl-L-leucyl-glycine nitrile
-
-
N-2,4,5-trifluorobenzoyl-L-leucyl-glycine nitrile
-
-
N-2,4-difluorobenzoyl-L-leucyl-glycine nitrile
-
-
N-2,5-difluorobenzoyl-L-leucyl-glycine nitrile
-
-
N-2,6-difluorobenzoyl-L-leucyl-glycine nitrile
-
-
N-2-fluorobenzoyl-L-alanine-beta-alanine nitrile
-
-
N-2-fluorobenzoyl-L-alanine-glycine nitrile
-
-
N-2-fluorobenzoyl-L-leucine-beta-alanine benzyl ester
-
-
N-2-fluorobenzoyl-L-leucine-beta-alanine nitrile
-
-
N-2-fluorobenzoyl-L-leucine-gamma-aminobutyric acid benzyl ester
-
-
N-2-fluorobenzoyl-L-leucyl-glycine benzyl ester
-
-
N-2-fluorobenzoyl-L-leucyl-glycine nitrile
-
-
N-2-trifluoromethylbenzoyl-L-leucyl-glycine nitrile
-
-
N-3,4,5-trifluorobenzoyl-L-leucyl-glycine nitrile
-
-
N-3,4-difluorobenzoyl-L-leucyl-glycine nitrile
-
-
N-3,5-difluorobenzoyl-L-leucyl-glycine nitrile
-
-
N-3-fluorobenzoyl-L-leucine-beta-alanine benzyl ester
-
-
N-3-fluorobenzoyl-L-leucine-gamma-aminobutyric acid benzyl ester
-
-
N-3-fluorobenzoyl-L-leucyl-glycine benzyl ester
-
-
N-3-fluorobenzoyl-L-leucyl-glycine nitrile
-
-
N-3-trifluoromethylbenzoyl-L-leucyl-glycine nitrile
-
-
N-4-fluorobenzoyl-L-leucine-beta-alanine benzyl ester
-
-
N-4-fluorobenzoyl-L-leucine-beta-alanine nitrile
-
-
N-4-fluorobenzoyl-L-leucine-gamma-aminobutyric acid benzyl ester
-
-
N-4-fluorobenzoyl-L-leucyl-glycine benzyl ester
-
-
N-4-fluorobenzoyl-L-leucyl-glycine nitrile
-
-
N-4-trifluoromethylbenzoyl-L-leucyl-glycine nitrile
-
-
N-benzoyl-L-leucyl-glycine
-
-
N-benzyloxycarbonylphenylalanyl-t-butyl-tryrosyl diazomethylketone
-
40.6% inhibition at 0.05 mM
N-cinnamoyl-L-leucyl-glycine nitrile
-
-
N-pentafluorobenzoyl-L-alanine-beta-alanine nitrile
-
-
N-pentafluorobenzoyl-L-alanine-glycine nitrile
-
-
N-pentafluorobenzoyl-L-leucine-beta-alanine benzyl ester
-
-
N-pentafluorobenzoyl-L-leucine-gamma-aminobutyric acid benzyl ester
-
-
N-pentafluorobenzoyl-L-leucyl-glycine benzyl ester
-
-
additional information
-
cathepsin L2 is completely inactivated by 4 mM tetranitromethane, cathepsin L1 is not inactivated
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
dithiothreitol
DTT
-
the action of FheCL1 is enhanced by glutathione, the major reducing agent found in red blood cells. In the presence of DTT, GSH and L-cysteine FheCL1 exhibited similar activation curves with maximal enzyme activity observed in the presence of each reducing agent at a concentration of 0.1 to 1.0 mM
glutathione
-
activitation, with maximum activity around 0.1-1 mM; the action of FheCL1 is enhanced by glutathione, the major reducing agent found in red blood cells. In the presence of DTT, GSH and L-cysteine FheCL1 exhibited similar activation curves with maximal enzyme activity observed in the presence of each reducing agent at a concentration of 0.1 to 1.0 mM
L-Cys
-
the action of FheCL1 is enhanced by glutathione, the major reducing agent found in red blood cells. In the presence of DTT, GSH and L-cysteine FheCL1 exhibited similar activation curves with maximal enzyme activity observed in the presence of each reducing agent at a concentration of 0.1 to 1.0 mM
L-cysteine
-
activitation, with maximum activity around 0.1-1 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0093
benzoyl-Phe-Val-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.0093
benzoyl-Phe-Val-Arg-7-amido-4-methylcoumarin
-
pH 7.0, 37°C
0.0204
benzyloxycarbonyl-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.0204
benzyloxycarbonyl-Arg-7-amido-4-methylcoumarin
-
pH 7.0, 37°C
0.0656
benzyloxycarbonyl-Arg-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.0656
benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin
-
pH 7.0, 37°C
0.38 - 4.35
benzyloxycarbonyl-L-Leu-L-Arg-4-methylcoumarinyl-7-amide
8.16 - 24.18
benzyloxycarbonyl-L-Phel-L-Arg-4-methylcoumarinyl-7-amide
48.41 - 191.2
benzyloxycarbonyl-L-Pro-L-Arg-4-methylcoumarinyl-7-amide
0.0036 - 0.0044
benzyloxycarbonyl-Leu-Arg-7-amido-4-methylcoumarin
0.0147
benzyloxycarbonyl-Phe-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.003 - 0.0242
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
0.1819 - 0.1912
benzyloxycarbonyl-Pro-Arg-7-amido-4-methylcoumarin
0.0054
H-Leu-Val-Tyr-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.0054
H-Leu-Val-Tyr-7-amido-4-methylcoumarin
-
pH 7.0, 37°C
0.0653
succinyl-Ala-Phe-Lys-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.0653
succinyl-Ala-Phe-Lys-7-amido-4-methylcoumarin
-
pH 7.0, 37°C
0.0385
succinyl-Leu-Leu-Val-Tyr-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.0385
succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin
-
pH 7.0, 37°C
0.0347
t-butyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin
-
pH 7.0, 37°C
0.0436
t-butyloxycarbonyl-Val-Pro-Arg-7-amido-4-methylcoumarin
-
pH 7.0, 37°C
10.43 - 21.57
tert-butoxycarbonyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
0.0347
tert-butyloxycarbonyl-Val-Leu-Lys-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.0436
tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.026 - 20.35
tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
0.026
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
-
pH 7.0, 37°C
0.1069
tosyl-Gly-Pro-Lys-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.1069
tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin
-
pH 7.0, 37°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.03
benzoyl-Phe-Val-Arg-4-methylcoumarinyl-7-amide
Fasciola hepatica
-
pH 7.0, 37°C, cathepsin L1
0.03
benzoyl-Phe-Val-Arg-7-amido-4-methylcoumarin
Fasciola hepatica
-
pH 7.0, 37°C
0.04
benzyloxycarbonyl-Arg-4-methylcoumarinyl-7-amide
Fasciola hepatica
-
pH 7.0, 37°C, cathepsin L1
0.04
benzyloxycarbonyl-Arg-7-amido-4-methylcoumarin
Fasciola hepatica
-
pH 7.0, 37°C
0.002
benzyloxycarbonyl-Arg-Arg-4-methylcoumarinyl-7-amide
Fasciola hepatica
-
pH 7.0, 37°C, cathepsin L1
0.002
benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin
Fasciola hepatica
-
pH 7.0, 37°C
1.73 - 36.52
benzyloxycarbonyl-L-Leu-L-Arg-4-methylcoumarinyl-7-amide
3.58 - 29.6
benzyloxycarbonyl-L-Phe-L-Arg-4-methylcoumarinyl-7-amide
0.122 - 1.03
benzyloxycarbonyl-L-Pro-L-Arg-4-methylcoumarinyl-7-amide
11 - 36.5
benzyloxycarbonyl-Leu-Arg-7-amido-4-methylcoumarin
1.08
benzyloxycarbonyl-Phe-Arg-4-methylcoumarinyl-7-amide
Fasciola hepatica
-
pH 7.0, 37°C, cathepsin L1
1.08 - 24.7
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
1 - 1.7
benzyloxycarbonyl-Pro-Arg-7-amido-4-methylcoumarin
0.02
H-Leu-Val-Tyr-4-methylcoumarinyl-7-amide
Fasciola hepatica
-
pH 7.0, 37°C, cathepsin L1
0.02
H-Leu-Val-Tyr-7-amido-4-methylcoumarin
Fasciola hepatica
-
pH 7.0, 37°C
0.05
succinyl-Ala-Phe-Lys-4-methylcoumarinyl-7-amide
Fasciola hepatica
-
pH 7.0, 37°C, cathepsin L1
0.05
succinyl-Ala-Phe-Lys-7-amido-4-methylcoumarin
Fasciola hepatica
-
pH 7.0, 37°C
0.01
succinyl-Leu-Leu-Val-Tyr-4-methylcoumarinyl-7-amide
Fasciola hepatica
-
pH 7.0, 37°C, cathepsin L1
0.01
succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin
Fasciola hepatica
-
pH 7.0, 37°C
7.9
t-butyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin
Fasciola hepatica
-
pH 7.0, 37°C
0.02
t-butyloxycarbonyl-Val-Pro-Arg-7-amido-4-methylcoumarin
Fasciola hepatica
-
pH 7.0, 37°C
0.2 - 0.93
tert-butoxycarbonyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
7.9
tert-butyloxycarbonyl-Val-Leu-Lys-4-methylcoumarinyl-7-amide
Fasciola hepatica
-
pH 7.0, 37°C, cathepsin L1
0.02
tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumarinyl-7-amide
Fasciola hepatica
-
pH 7.0, 37°C, cathepsin L1
0.03 - 0.36
tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
0.03
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
Fasciola hepatica
-
pH 7.0, 37°C
0.03
tosyl-Gly-Pro-Lys-4-methylcoumarinyl-7-amide
Fasciola hepatica
-
pH 7.0, 37°C, cathepsin L1
0.03
tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin
Fasciola hepatica
-
pH 7.0, 37°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0000154 - 0.001125
benzyloxycarbonyl-Phe-Ala-diazomethyl ketone
0.0000108 - 0.000116
cathepsin K inhibitor II
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.003
Benzyloxycarbonyl-Phe-Ala-CHN2
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0044
N-2,3,4,5,6-pentafluorobenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0034
N-2,3,4,5-tetrafluorobenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0041
N-2,3,4-trifluorobenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.006
N-2,3,6-trifluorobenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0034
N-2,3-difluorobenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0045
N-2,4,5-trifluorobenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0055
N-2,4-difluorobenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0031
N-2,5-difluorobenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0037
N-2,6-difluorobenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.1
N-2-fluorobenzoyl-L-alanine-beta-alanine nitrile
Fasciola hepatica
-
IC50 above 0.1 mM, in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0876
N-2-fluorobenzoyl-L-alanine-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0294
N-2-fluorobenzoyl-L-leucine-beta-alanine benzyl ester
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0243
N-2-fluorobenzoyl-L-leucine-beta-alanine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0153
N-2-fluorobenzoyl-L-leucine-gamma-aminobutyric acid benzyl ester
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.1
N-2-fluorobenzoyl-L-leucyl-glycine benzyl ester
Fasciola hepatica
-
IC50 above 0.1 mM, in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.003
N-2-fluorobenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0041
N-2-trifluoromethylbenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0034
N-3,4,5-trifluorobenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0031
N-3,4-difluorobenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.003
N-3,5-difluorobenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0248
N-3-fluorobenzoyl-L-leucine-beta-alanine benzyl ester
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0401
N-3-fluorobenzoyl-L-leucine-gamma-aminobutyric acid benzyl ester
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.1
N-3-fluorobenzoyl-L-leucyl-glycine benzyl ester
Fasciola hepatica
-
IC50 above 0.1 mM, in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0031
N-3-fluorobenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0091
N-3-trifluoromethylbenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0623
N-4-fluorobenzoyl-L-leucine-beta-alanine benzyl ester
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0216
N-4-fluorobenzoyl-L-leucine-beta-alanine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0193
N-4-fluorobenzoyl-L-leucine-gamma-aminobutyric acid benzyl ester
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0434
N-4-fluorobenzoyl-L-leucyl-glycine benzyl ester
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0028
N-4-fluorobenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0032
N-4-trifluoromethylbenzoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.01
N-benzoyl-L-leucyl-glycine
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.011
N-cinnamoyl-L-leucyl-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.1
N-pentafluorobenzoyl-L-alanine-beta-alanine nitrile
Fasciola hepatica
-
IC50 above 0.1 mM, in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0683
N-pentafluorobenzoyl-L-alanine-glycine nitrile
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0577
N-pentafluorobenzoyl-L-leucine-beta-alanine benzyl ester
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0214
N-pentafluorobenzoyl-L-leucine-gamma-aminobutyric acid benzyl ester
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
0.0395
N-pentafluorobenzoyl-L-leucyl-glycine benzyl ester
Fasciola hepatica
-
in 0.1 M sodium acetate, at 37°C, pH not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
16.2
-
substrate: tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.2
-
; maximal activity between pH 5.5–7.0 with a peak at pH 6.2, substrate: benzyloxycarbonyl-L-Phe-L-Arg-4-methylcoumarinyl-7-amide
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 9
-
the enzyme has the capacity to cleave substrates over a wide pH range
5.5 - 7
-
substrate ovalbumin
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.6
-
calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
highest expression
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
-
epithelial cells, inactive proform procathepsin L1 is packaged in secretory vesicles lining the parasite gut, activation of the proform takes place following secretion of the protease into the gut lumen
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
23000
-
x * 23000, active enzyme, SDS-PAGE; x * 23000, mature enzyme, SDS-PAGE
24500
-
sequence analysis
29000
sequence data
30000
-
x * 30000, recombinant enzyme, SDS-PAGE
36950
recombinant protein, calculated from amino acid sequence
37130
-
x * 37130, calculated from amino acid sequence
38000
-
x * 25000, mature enzyme, x * 38000, zymogen, SDS-PAGE
60000
-
x * 60000, mature GST- and His6-tagged enzyme, SDS-PAGE; x * 60000, recombinant enzyme with GST and His6 tags, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
-
the enzyme contains three potential glycosylation sites at residues 18 (N-A-S), 38 (N-S-S), and 99 (N-A-A)
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
the 1.4 A three-dimensional structure of the FheCL1 was determined by x-ray crystallography
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.5
-
37°C, enzyme activity is not completely lost until day three
694804
3
-
37°C, 10 days, 1 mM DTT, 95% loss of activity, mature FheCL1
694804
4.5 - 7.5
-
zymogen is stable within this range
694804
4.5
-
37°C, 10 days, 1 mM DTT, 55% loss of activity, mature FheCL1; retains 45% of activity when incubated at 37°C and pH 4.5 for 10 days
694804
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
retains 45% of activity when incubated at 37°C and pH 4.5 for 10 days
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ethanol
-
no inactivation of cathepsin L1 or cathepsin L2
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
combined purification of cathepsin L1 and cathepsin L2
-
from 1.5% agarose gel
gel filtration, proenzyme purified by affinity Ni-NTA-agarose chromatography
-
in the presence of 1% Triton X-100 by Ni-NTA affinity column chromatography and Sephacryl S-100 gel filtration; Ni-NTA affinity column chromatography and gel filtration
-
Ni-NTA affinity column chromatography
Ni-Sepharose resin column chromatography, gel filtration
Ni2+-NTA affinity column chromatography
-
nickel affinity column chromatography
-
recombinant zymogen of cathepsin L1
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a 25 kDa leucine aminopeptidase-cathepsin L1 chimeric protein is expressed from the pET15b plasmid in the Escherichia coli strain Rosetta (DE3)
cloned from a Turkish isolate
expressed as a fusion protein with GST and His6 tags in Escherichia coli Rosetta (DE3) pLysS cells; expressed in Escherichia coli Rosetta (DE3) pLysS cells
-
expressed in Escherichia coli BL21(DE) cells
-
expressed in Escherichia coli BL21-SI cells
expressed in Pichia pastoris
-
expressed in Saccharomyces cerevisiae
expression in Escherichia coli
-
expression in Pichia pastoris
-
expression of pro-form in yeast
-
expression of recombinant proenzyme in Pichia pastoris
-
wild-type and variant enzymes are recombinantly expressed in Pichia pastoris
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the expression level of CL1 mRNA is significantly down-regulated at 12 h and 24 h in digestive gland after Vibrio alginolyticus stimulation
the expression level of CL1 mRNA is significantly up-regulated at 4 h and 8 h in digestive gland after Vibrio alginolyticus stimulation
under starvation conditions, cathepsin L1 expression is decreased at 30 days
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C25G
-
inactive zymogen since the active site Cys is replaced by a Gly
C26G
-
active site variant FheproCL1C26G cannot autocatalytically process. It is susceptible to trans-processing at a Leu12-Ser11/His10 sequence by preactivated FheCL1
L12P
-
the autoactivation of the variant enzyme FheproCL1L12P is very slow but is increased 40fold in the presence of FheCL2
L12P/C26G
-
active site variant FheproCL1L12P/C26G cannot autocatalytically process. It is not susceptible to trans-processing at a Leu12-Ser11/His10 sequence by preactivated FheCL1. Another Fasciola hepatica secreted protease FheCL2, which, unlike FheCL1, can readily accept proline in the S2 subsite of its active site, can trans-process the double variant FheproCL1L12P/C26G by cleavage at the Pro12-Ser11/His10 sequence
L205A
-
mutation of FheCL1 markedly alters the activity profile from wild type enzyme. This variant exhibits a broader substrate specificity by accepting Phe, Trp, and Tyr at P2, residues that are not accepted by wild type FheCL1. kcat/Km for benzyloxycarbonyl-L-Phe-L-Arg-4-methylcoumarinyl-7-amide is 1.5fold higher than wild-type value, kcat/Km for benzyloxycarbonyl-L-Leu-L-Arg-4-methylcoumarinyl-7-amide is 2.5fold lower than wild-type value, kcat/Km for benzyloxycarbonyl-Pro-L-Arg-4-methylcoumarinyl-7-amide is 2.2fold lower than wild-type value, kcat/Km for tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide is 6.6fold lower than wild-type value, kcat/Km for tert-butoxycarbonyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide is 5fold than wild-type value
L209A
-
Ki value for benzyloxycarbonyl-Phe-Ala-diazomethyl ketone is 73fold lower than value for wild-type FheCL1
L67Y
-
no significant change in the P2 preference to wild type FheCL1. This substitution does not alter the activity of the enzyme toward Pro in the P2 position. kcat/Km for benzyloxycarbonyl-L-Phe-L-Arg-4-methylcoumarinyl-7-amide is 2.3fold lower than wild-type value, kcat/Km for benzyloxycarbonyl-L-Leu-L-Arg-4-methylcoumarinyl-7-amide is 1.8fold lower than wild-type value, kcat/Km for benzyloxycarbonyl-Pro-L-Arg-4-methylcoumarinyl-7-amide is 1.2fold lower than wild-type value, kcat/Km for tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide is similar to wild-type value, kcat/Km for tert-butoxycarbonyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide is 1.8fold higher than wild-type value. Ki value for benzyloxycarbonyl-Phe-Ala-diazomethyl ketone is 12fold lower than wild-type value
additional information
-
screening of cathepsin L1/cathepsin L2 mimotopes and use of an M13 phage random 12-mers peptide library to evaluate their immunogenicity in sheep
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
medicine