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Information on EC 3.4.22.B49 - cathepsin L1 and Organism(s) Fasciola hepatica and UniProt Accession Q24940
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3.4.22.B49 cathepsin L1Specify your search results
Word Map
- 3.4.22.B49
- cathepsins
- fasciola
- hepatica
- fluke
-
fasciolosis
-
metacercariae
-
helminth
- gigantica
- medicine
-
excysted
-
trematode
-
excretory-secretory
-
immunodiagnosis
- agriculture
-
mimotopes
- diagnostics
The taxonomic range for the selected organisms is: Fasciola hepatica
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
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
Synonyms
cathepsin l1, ctsl1, fhcl1, fhecl1, catl1, fgcatl1h, cgctsl1, cpfhw, cathepsin l1h, da-ctsl1, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
cathepsin L1
cathepsin L1 proteinase
-
-
-
-
CL1
FhCL1
FheCL1
cathepsin L1
-
-
-
-
CL1
-
-
-
-
FheCL1
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
60616-82-2
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
benzyloxycarbonyl-L-Leu-L-Arg-4-methylcoumarinyl-7-amide + H2O
benzyloxycarbonyl-L-Leu-L-Arg + 7-amino-4-methylcoumarin
FheCL1
-
-
?
benzyloxycarbonyl-L-Phe-L-Arg-4-methylcoumarinyl-7-amide + H2O
benzyloxycarbonyl-L-Phe-L-Arg + 7-amino-4-methylcoumarin
FheCL1
-
-
?
benzyloxycarbonyl-L-Pro-L-Arg-4-methylcoumarinyl-7-amide + H2O
benzyloxycarbonyl-L-Pro-L-Arg + 7-amino-4-methylcoumarin
FheCL1
-
-
?
Collagen + H2O
?
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
-
-
?
Hemoglobin + H2O
?
FhCL1 cleaves substrates with hydrophobic residues (Phe and Leu) in the P2 position with catalytic rates (kcat/Km) that are 25- and eightfold greater, respectively, than FhCL2. In comparison to human cathepsin L, which can accommodate a wide range of amino acids in the S2 subsite, the S2 subsite of FhCL1 is restricted. Hydrophobic residues are most susceptible to cleavage, in the order Leu > Val > Ala > Phe. Together, these four residues make up about 42% of the hemoglobin molecule and, therefore, it seems that FhCL1 has been specifically adapted to degrade the host substrate, which it exploits as nutrient. Substrates with proline in the P2 position, which are good substrates for FhCL2 are poorly cleaved by FhCL1 (and not at all by human cathepsin L)
-
-
?
tert-butoxycarbonyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide + H2O
tert-butoxycarbonyl-Gly-Pro-Arg + 7-amino-4-methylcoumarin
FheCL1
-
-
?
tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide + H2O
tosyl-Gly-Pro-Arg + 7-amino-4-methylcoumarin
FheCL1
-
-
?
benzoyl-Phe-Val-Arg-4-methylcoumarinyl-7-amide + H2O
benzoyl-Phe-Val-Arg + 7-amino-4-methylcoumarin
-
cathepsin L1
-
-
?
benzoyl-Phe-Val-Arg-7-amido-4-methylcoumarin + H2O
benzoyl-Phe-Val-Arg + 7-amino-4-methylcoumarin
-
-
-
-
?
benzyloxycarbonyl-Arg-4-methylcoumarinyl-7-amide + H2O
benzyloxycarbonyl-Arg + 7-amino-4-methylcoumarin
-
cathepsin L1
-
-
?
benzyloxycarbonyl-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Arg + 7-amino-4-methylcoumarin
-
-
-
-
?
benzyloxycarbonyl-Arg-Arg-4-methylcoumarinyl-7-amide + H2O
benzyloxycarbonyl-Arg-Arg + 7-amino-4-methylcoumarin
-
cathepsin L1
-
-
?
benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Arg-Arg + 7-amino-4-methylcoumarin
-
-
-
-
?
benzyloxycarbonyl-L-Phe-L-Arg-4-methylcoumarinyl-7-amide + H2O
benzyloxycarbonyl-L-Phe-L-Arg + 7-amino-4-methylcoumarin
-
FheCL1
-
-
?
benzyloxycarbonyl-L-phenylalanyl-L-arginine 4-methylcoumarinyl-7-amide + H2O
?
-
-
-
-
?
benzyloxycarbonyl-Leu-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Leu-Arg + 7-amino-4-methylcoumarin
-
-
-
?
benzyloxycarbonyl-Phe-Arg-4-methylcoumarinyl-7-amide + H2O
benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
-
cleaved by cathepsin L2 with much greater affinity than by cathepsin L1
-
-
?
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
benzyloxycarbonyl-Phe-Phe-Arg-7-amido-4-methylcoumarin + H2O
?
-
renatured CPFhW
-
-
?
benzyloxycarbonyl-Pro-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Pro-Arg + 7-amino-4-methylcoumarin
-
-
-
?
Fibrinogen + H2O
?
enzyme FhCL3 is capable of degradation of the fibrinogen alpha-chain, beta-chain, and gamma-chain
-
-
?
H-Leu-Val-Tyr-4-methylcoumarinyl-7-amide + H2O
H-Leu-Val-Tyr + 7-amino-4-methylcoumarin
-
cleaved by cathepsin L2 with much greater affinity than by cathepsin L1
-
-
?
H-Leu-Val-Tyr-7-amido-4-methylcoumarin + H2O
H-Leu-Val-Tyr + 7-amino-4-methylcoumarin
-
-
-
-
?
human IgG + H2O
?
-
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
-
cleaved by cathepsin L2 with much greater affinity than by cathepsin L1
-
-
?
procathepsin L1 + H2O
?
-
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
-
cathepsin L1
-
-
?
succinyl-Ala-Phe-Lys-7-amido-4-methylcoumarin + H2O
tosyl-Ala-Phe-Lys + 7-amino-4-methylcoumarin
-
-
-
-
?
succinyl-Leu-Leu-Val-Tyr-4-methylcoumarinyl-7-amide + H2O
succinyl-Leu-Leu-Val-Tyr + 7-amino-4-methylcoumarin
-
cathepsin L1
-
-
?
succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin + H2O
succinyl-Leu-Leu-Val-Tyr + 7-amino-4-methylcoumarin
-
-
-
-
?
t-butyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin + H2O
t-butyloxycarbonyl-Val-Leu-Lys + 7-amino-4-methylcoumarin
-
-
-
-
?
t-butyloxycarbonyl-Val-Pro-Arg-7-amido-4-methylcoumarin + H2O
t-butyloxycarbonyl-Val-Pro-Arg + 7-amino-4-methylcoumarin
-
cleaved by cathepsin L2 with much greater affinity than by cathepsin L1
-
-
?
tert-butyloxycarbonyl-Val-Leu-Lys-4-methylcoumarinyl-7-amide + H2O
tert-butyloxycarbonyl-Val-Leu-Lys + 7-amino-4-methylcoumarin
-
cathepsin L1
-
-
?
tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumarinyl-7-amide + H2O
tert-butyloxycarbonyl-Val-Pro-Arg + 7-amino-4-methylcoumarin
-
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
-
cathepsin L1
-
-
?
tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide + H2O
tosyl-Gly-Pro-Arg + 7-amino-4-methylcoumarin
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin + H2O
tosyl-Gly-Pro-Arg + 7-amino-4-methylcoumarin
-
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
-
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
-
cleaved by cathepsin L2 with much greater affinity than by cathepsin L1
-
-
?
Z-Phe-Arg-OMe + SerNH2
Z-Phe-Arg-Ser-NH2 + methanol
-
peptide synthesis
-
-
?
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
-
-
-
-
?
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
-
-
-
?
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
-
cleaved by cathepsin L2 with much greater affinity than by cathepsin L1
-
-
?
Hemoglobin + H2O
?
-
Fasciola hepatica secretes cathepsin L cysteine proteases to invade its host, migrate through tissues and digest hemoglobin, its main source of amino acids. FheCL1 can degrade hemoglobin to small peptides, predominantly of 414 residues, but cannot release free amino acids. It is suggested that hemoglobin degradation is not completed in the gut lumen but the resulting peptides are absorbed by the gut epithelial cells for further processing by intracellular di- and amino-peptidases to free amino acids that are distributed through the parasite tissue for protein anabolism. The action of FheCL1 is enhanced by glutathione, the major reducing agent found in red blood cells
-
-
?
Hemoglobin + H2O
?
-
FheCL1 can degrade hemoglobin to small peptides, predominantly of 414 residues, but cannot release free amino acids. FheCL1 can not cleave its natural substrate hemoglobin in the pH range pH 5.5 and pH 7.0. Digestion occurs only at or below pH 4.5, which coincides with pH-induced dissociation of the hemoglobin tetramer. The acidic pH of the parasite relaxes the hemoglobin structure, making it susceptible to proteolysis by FheCL1. The P1 position could be occupied by many amino acids but most preferentially Leu. FheCL1 preferentially cleaves bonds where the P2 position is occupied with hydrophobic residues (in order of decreasing efficiency: Leu, Val, Ala, Phe) and is observed for the digestion of both hemoglobin-alpha and hemoglobin-beta
-
-
?
Hemoglobin + H2O
?
-
enzyme does not cleave its natural substrate hemoglobin in the pH range pH 5.5 and pH 7.0. Digestion occurs only at pH 4.5, which coincides with pH-induced dissociation of the hemoglobin tetramer
degradation to small peptides of 4-14 residues, no release of free amino acids
-
?
ovalbumin + H2O
?
-
FheCL1 can degrade ovalbumin from pH 3.5 to pH 8.0
-
-
?
tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide + H2O
tosyl-Gly-Pro-Arg + 7-amino-4-methylcoumarin
-
-
-
-
?
tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide + H2O
tosyl-Gly-Pro-Arg + 7-amino-4-methylcoumarin
-
cleaved by cathepsin L2 with much greater affinity than by cathepsin L1
-
-
?
additional information
?
-
wild-type FheCL1 shows clear preference for Arg at P1. Other residues accommodated in this position including Lys, Glu, Thr, and Met are all cleaved at similar relative rates to that observed for human cathepsin L and cathepsin K. Similar results are obtained for the variants FheCL1 L67Y and FheCL1 L205A. FheCL1 shows distinct preference for hydrophobic amino acids in the P2, Leu is favored. FheCL1 and FheCL2 are similar to cathepsin K with regard to their preference for a P2 Leu over Phe (human cathepsin L has a preference for P2 Phe over Leu). Both enzymes can accommodate Pro in the P2 position, but this is more readily accepted by FheCL2 compared with FheCL1. Neither enzyme, however, cleaves substrates with this residue in the P2 position as readily as human cathepsin K
-
-
?
additional information
?
-
-
wild-type FheCL1 shows clear preference for Arg at P1. Other residues accommodated in this position including Lys, Glu, Thr, and Met are all cleaved at similar relative rates to that observed for human cathepsin L and cathepsin K. Similar results are obtained for the variants FheCL1 L67Y and FheCL1 L205A. FheCL1 shows distinct preference for hydrophobic amino acids in the P2, Leu is favored. FheCL1 and FheCL2 are similar to cathepsin K with regard to their preference for a P2 Leu over Phe (human cathepsin L has a preference for P2 Phe over Leu). Both enzymes can accommodate Pro in the P2 position, but this is more readily accepted by FheCL2 compared with FheCL1. Neither enzyme, however, cleaves substrates with this residue in the P2 position as readily as human cathepsin K
-
-
?
additional information
?
-
-
the enzyme mediates kinin release from high molecular weight kininigen
-
-
?
additional information
?
-
-
the enzyme facilitates the penetration of the parasite though the tissue of its host, and also participates in functions such as feeding and immune evasion
-
-
?
additional information
?
-
-
cathepsin L1 and cathepsin L2 proteinases may be the prime mechanism by which the parasite penetrates tissue
-
-
?
additional information
?
-
-
renatured enzyme shows no activity with benzyloxycarbonyl-Phe-Arg-beta-naphthylamide
-
-
?
additional information
?
-
-
enzyme preferentially cleaves bonds where the P2 position is occupied with hydrophobic residues, this preference follows the order Leu>Val>Ala>Phe, and is observed for the digestion of both hemoglobin-alpha and hemoglobin-beta
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
Hemoglobin + H2O
?
-
Fasciola hepatica secretes cathepsin L cysteine proteases to invade its host, migrate through tissues and digest hemoglobin, its main source of amino acids. FheCL1 can degrade hemoglobin to small peptides, predominantly of 414 residues, but cannot release free amino acids. It is suggested that hemoglobin degradation is not completed in the gut lumen but the resulting peptides are absorbed by the gut epithelial cells for further processing by intracellular di- and amino-peptidases to free amino acids that are distributed through the parasite tissue for protein anabolism. The action of FheCL1 is enhanced by glutathione, the major reducing agent found in red blood cells
-
-
?
Hemoglobin + H2O
?
-
enzyme does not cleave its natural substrate hemoglobin in the pH range pH 5.5 and pH 7.0. Digestion occurs only at pH 4.5, which coincides with pH-induced dissociation of the hemoglobin tetramer
degradation to small peptides of 4-14 residues, no release of free amino acids
-
?
additional information
?
-
-
the enzyme facilitates the penetration of the parasite though the tissue of its host, and also participates in functions such as feeding and immune evasion
-
-
?
additional information
?
-
-
cathepsin L1 and cathepsin L2 proteinases may be the prime mechanism by which the parasite penetrates tissue
-
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
cathepsin L2 is completely inactivated by 4 mM tetranitromethane, cathepsin L1 is not inactivated
-
additional information
the enzyme's fibrinolytic activity is inhibited by plasma. Addition of GSH to plasma cannot counteract the inhibitory effect of plasma components for FhCL1
-
additional information
-
the enzyme's fibrinolytic activity is inhibited by plasma. Addition of GSH to plasma cannot counteract the inhibitory effect of plasma components for FhCL1
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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
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
dithiothreitol
-
the cathepsins L needed the presence of dithiothreitol to digest IgG1, IgG2, and IgG4 whereas IgG3 was identically cleaved under both reducing and nonreducing conditions
glutathione
-
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
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0093
benzoyl-Phe-Val-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.0204
benzyloxycarbonyl-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.0656
benzyloxycarbonyl-Arg-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.0147
benzyloxycarbonyl-Phe-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.0054
H-Leu-Val-Tyr-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.0653
succinyl-Ala-Phe-Lys-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.0385
succinyl-Leu-Leu-Val-Tyr-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
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
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
tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.1069
tosyl-Gly-Pro-Lys-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.38
benzyloxycarbonyl-L-Leu-L-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L67Y FheCL1
2.75
benzyloxycarbonyl-L-Leu-L-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L205A FheCL1
4.35
benzyloxycarbonyl-L-Leu-L-Arg-4-methylcoumarinyl-7-amide
wild-type enzyme FheCL1
8.16
benzyloxycarbonyl-L-Phel-L-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L67Y FheCL1
19.21
benzyloxycarbonyl-L-Phel-L-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L205A FheCL1
24.18
benzyloxycarbonyl-L-Phel-L-Arg-4-methylcoumarinyl-7-amide
wild-type enzyme FheCL1
48.41
benzyloxycarbonyl-L-Pro-L-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L205A FheCL1
137
benzyloxycarbonyl-L-Pro-L-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L67Y FheCL1
191.2
benzyloxycarbonyl-L-Pro-L-Arg-4-methylcoumarinyl-7-amide
wild-type enzyme FheCL1
10.43
tert-butoxycarbonyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
wild-type enzyme FheCL1
11.13
tert-butoxycarbonyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L67Y FheCL1
21.57
tert-butoxycarbonyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L205A FheCL1
6.96
tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L67Y FheCL1
10.02
tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
wild-type enzyme FheCL1
20.35
tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L205A FheCL1
0.0036
benzyloxycarbonyl-Leu-Arg-7-amido-4-methylcoumarin
37°C, pH 7.3
0.0044
benzyloxycarbonyl-Leu-Arg-7-amido-4-methylcoumarin
37°C, pH 5.5
0.003
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
37°C, pH 7.3
0.0147
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
-
pH 7.0, 37°C
0.0242
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
37°C, pH 5.5
0.1819
benzyloxycarbonyl-Pro-Arg-7-amido-4-methylcoumarin
37°C, pH 7.3
0.1912
benzyloxycarbonyl-Pro-Arg-7-amido-4-methylcoumarin
37°C, pH 5.5
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.03
benzoyl-Phe-Val-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.04
benzyloxycarbonyl-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.002
benzyloxycarbonyl-Arg-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
1.08
benzyloxycarbonyl-Phe-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.05
succinyl-Ala-Phe-Lys-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.01
succinyl-Leu-Leu-Val-Tyr-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.02
t-butyloxycarbonyl-Val-Pro-Arg-7-amido-4-methylcoumarin
-
pH 7.0, 37°C
7.9
tert-butyloxycarbonyl-Val-Leu-Lys-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.02
tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.03
tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
0.03
tosyl-Gly-Pro-Lys-4-methylcoumarinyl-7-amide
-
pH 7.0, 37°C, cathepsin L1
1.73
benzyloxycarbonyl-L-Leu-L-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L67Y FheCL1
9.15
benzyloxycarbonyl-L-Leu-L-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L205A FheCL1
36.52
benzyloxycarbonyl-L-Leu-L-Arg-4-methylcoumarinyl-7-amide
wild-type enzyme FheCL1
3.58
benzyloxycarbonyl-L-Phe-L-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L67YFheCL1
24.69
benzyloxycarbonyl-L-Phe-L-Arg-4-methylcoumarinyl-7-amide
wild-type enzyme, FheCL1
29.6
benzyloxycarbonyl-L-Phe-L-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L205A FheCL1
0.122
benzyloxycarbonyl-L-Pro-L-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L205A FheCL1
0.62
benzyloxycarbonyl-L-Pro-L-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L67Y FheCL1
1.03
benzyloxycarbonyl-L-Pro-L-Arg-4-methylcoumarinyl-7-amide
wild-type enzyme FheCL1
0.2
tert-butoxycarbonyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L205A FheCL1
0.48
tert-butoxycarbonyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
wild-type enzyme FheCL1
0.93
tert-butoxycarbonyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L67Y FheCL1
0.113
tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L205A FheCL1
0.26
tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
mutant enzyme L67Y FheCL1
0.36
tosyl-Gly-Pro-Arg-4-methylcoumarinyl-7-amide
wild-type enzyme FheCL1
36.5
benzyloxycarbonyl-Leu-Arg-7-amido-4-methylcoumarin
37°C, pH 5.5
24.7
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
37°C, pH 5.5
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0000154
benzyloxycarbonyl-Phe-Ala-diazomethyl ketone
mutant enzyme L209A, FheCL1
0.0000936
benzyloxycarbonyl-Phe-Ala-diazomethyl ketone
mutant enzyme L67Y, FheCL1
0.001125
benzyloxycarbonyl-Phe-Ala-diazomethyl ketone
wild-type enzyme, FheCL1
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
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
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.2
6.2
-
maximal activity between pH 5.57.0 with a peak at pH 6.2, substrate: benzyloxycarbonyl-L-Phe-L-Arg-4-methylcoumarinyl-7-amide
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
3 - 9
-
the enzyme has the capacity to cleave substrates over a wide pH range
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
additional information
CL1 belongs to a variety of cysteine proteases that are extensively expressed in the Fasciola species
-
the slightly acidic pH value of gut facilitates auto-activation of the inactive zymogen proCL1
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
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
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
adult flukes produce three different clades of cathepsin Ls: FhCL1, FhCL2, and FhCL5
physiological function
Although no direct anticoagulant effect of the peptidases is observed, cathepsin peptidases from Fasciola are able to degrade purified fibrinogen, with FhCL1 having the highest fibrinogenolytic activity. FhCL1 and FhCL2 also both efficiently degraded fibrin, but FhCL3 does not. FhCL1 has a larger fibrinogenolytic activity than FhCL2 and FhCL3 and is capable of degradation of the fibrinogen alpha-chain, beta-chain, and gamma-chain. FhCL2 andFhCL3 demonstrate only minor cleavage of the gamma-chain and slower cleavage of the alpha-chain and beta-chain compared to FhCL1
additional information
peptides QWKRMYNKEYNGADDEHRRNIWEENV and DKIDWRESGYVTELKDQGNC from the sequence of cathepsin L1 from Fasciola gigantica and the three-dimensional structure of cathepsin L1 from Fasciola hepatica, UniProt ID Q24940 and PDB ID 2o6x, are used for modelling of diagnostic peptides binding to cathepsinL1 antigen in fasciolosis, overview
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
x * 25000, about, recombinant chimeric LAP-CL1 mutant enzyme, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
proteolytic modification
FhCL1 is expressed as 37000 Da zymogen that autocatalytically processes at pH 4.5 to produce a 24500 Da mature enzyme
proteolytic modification
proteolytic modification
-
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. 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. The autoactivation of a variant enzyme with a single replacement, FheproCL1L12P, is very slow but is increased 40fold in the presence of FheCL2
proteolytic modification
-
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. 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. The autoactivation of a variant enzyme with a single replacement, FheproCL1L12P, is very slow but is increased 40fold in the presence of FheCL2
proteolytic modification
-
slightly acidic pH of the parasite gut facilitates the auto-catalytic activation of FheCL1 from its inactive proFheCL1 zymogen. This process is about 40fold faster at pH 4.5 than at pH 7.0. The zymogen proFheCL1 (38000 Da) is auto-catalytically processed at pH 4.5 by intermolecular cleavage and removal of the prosegment to release a fully mature and active enzyme (25000 Da)
proteolytic modification
-
auto-activation of the inactive zymogen proCL1 in the slightly acidic gut. The autoactivation process is 40fold faster at pH 4.5 than at pH 7.0. Molecular weight of zymogen is 38 kDa, of mature form 25 kDA, respectively
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
the 1.4 A three-dimensional structure of the FheCL1 was determined by x-ray crystallography
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
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
additional information
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
additional information
construction and evaluation of a chimeric LAP-CL1 mutant made from Fasciola hepatica leucine ainopeptidase and cathepsin L1. The protein contains the most antigenic sequences of LAP (amino acids 192-281, UniProt ID Q17TZ3) and CL1 (amino acids 173-309)
additional information
-
construction and evaluation of a chimeric LAP-CL1 mutant made from Fasciola hepatica leucine ainopeptidase and cathepsin L1. The protein contains the most antigenic sequences of LAP (amino acids 192-281, UniProt ID Q17TZ3) and CL1 (amino acids 173-309)
additional information
construction of a chimeric protein from leucine aminopeptidase (FhLAP) and cathepsin L1 (FhCL1) of Fasciola hepatica. The chimeric mutant protein with Quil A adjuvant is used for vaccination of Katahdin x East Friesian male sheep
additional information
-
construction of a chimeric protein from leucine aminopeptidase (FhLAP) and cathepsin L1 (FhCL1) of Fasciola hepatica. The chimeric mutant protein with Quil A adjuvant is used for vaccination of Katahdin x East Friesian male sheep
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4.5
4.5
-
retains 45% of activity when incubated at 37°C and pH 4.5 for 10 days
694804
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
gel filtration, proenzyme purified by affinity Ni-NTA-agarose chromatography
-
recombinant His-tagged chimeric mutant from Escherichia coli strain Rosetta (DE3) by nickel affinity chromatography and dialysis
recombinant solubilized and refolded His-tagged enzyme from Escherichia coli by nickel affinity chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
wild-type and variant enzymes are recombinantly expressed in Pichia pastoris
a 25 kDa leucine aminopeptidase-cathepsin L1 chimeric protein is expressed from the pET15b plasmid in the Escherichia coli strain Rosetta (DE3)
gene FhpCL1, DNA and amino acid sequence determination and analysis, sequence comparisons of rFhpCLs, recombinant expression of His-tagged enzyme in Escherichia coli
recombinant expression of His-tagged chimeric LAP-CL1 mutant in Escherichia coli strain Rosetta (DE3)
recombinant expression of His-tagged chimeric mutant from leucine aminopeptidase (FhLAP) and cathepsin L1 (FhCL1) of Fasciola hepatica in Escherichia coli strain Rosetta (DE3)
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged enzyme from Escherichia coli inclusion bodies, solubilization by 8 M urea, followed by dilution, dialysis, and ultrafiltration
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
diagnostics
design and synthesis of a new peptide derived from Fasciola gigantica cathepsin L1 with potential application in serodiagnosis of fascioliasis via ELISA, overview. Cathepsin L1 as antigen for serodiagnosis of animal fasciolosis
agriculture
diagnostics
analysis of the diagnostic values of the three different clades of cathepsin Ls, FhCL1, FhCL2, and FhCL5, from adult flukes in an ELISA, test of sera from sheep and cattle naturally infected with Fasciola hepatica, of cross-reactive antibodies, overview. For sheep sera, the sensitivity is 100% for the three rFhpCLs, while for cattle sera, the highest sensitivity is obtained using rFhpCL2 (97%), being equal for both rFhpCL1 and rFhpCL5 (87.9%), after adjusting cut-offs for maximum specificity
medicine
agriculture
-
screening of cathepsin L1/cathepsin L2 mimotopes and use of an M13 phage random 12-mers peptide library to evaluate their immunogenicity in sheep. Immunization of sheep with clones showing positive reactivity to rabbit cathepsin L1/L2 antiserum results in decrease in worm burdens after challenge. A significant reduction in worm size and burden is observed for those sheep immunized with clone 1. Animals receiving clone 20, show a significant reduction in egg output. Immunization induces a reduction of egg viability ranging from 58% to 82%. Vaccinated animals produce clone-specific antibodies which are boosted after challenge with metacercariae of Fasciola hepatica
agriculture
-
vaccination of rats with recombinant zymogen and subsequent challenge with fasciola hepatica metercercariae. Vaccination results in significantly smaller and fewer flukes than in controls
medicine
-
potential in the development of first generation liver fluke vaccines. Vaccine trials in both sheep and cattle with purified native FheCL1 and FheCL2 show that the enzymes can induce protection, ranging from 33-79%, to experimental challenge with metacercariae of Fasciola hepatica, and very potent anti-embryonation/hatch rate effects that would block parasite transmission
medicine
the cathepsin L1 gene may be used for DNA vaccination, recombinant protein derived from the gene can be used for serological diagnostics against Fasciola hepatica in Turkey
medicine
-
enteral vaccination of rats against Fasciola hepatica using recombinant cysteine proteinase (cathepsin L1). The mature CPFhW protein is the enzyme secreted by Fasciola hepatica with which the host organism has contact. This can explain why immunisation with this antigen resultes in best protection in challenged animals, especially in comparison to the immature form of the CPFhW protein. Substantial protection can be obtained when the antigen is given with food. Oral administration of the antigen does not lead to decreasing of the level of protection compared with intragastric administration
medicine
-
cattle can be protected against a natural infection of Fasciola hepatica by vaccination with recombinant cathepsin L1
medicine
a leucine aminopeptidase-cathepsin L1 chimeric protein may be used as a diagnostic tool for detection of antibodies against Fasciola hepatica in bovine sera and as an immunogen to induce protection against bovine fasciolosis
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Dowd, A.J.; Smith, A.M.; McGonigle, S.; Dalton, J.P.
Purification and characterization of a second cathepsin L proteinase secreted by parasitic trematode Fasciola hepatica
Eur. J. Biochem.
223
91-98
1994
Fasciola hepatica
Smith, A.M.; Dowd, A.J.; McGonigle, S.; Keegan, P.S.; Brennan, G.; Trudgett, A.; Dalton, J.P.
Purification of a cathepsin L-like proteinase secreted by adult Fasciola hepatica
Mol. Biochem. Parasitol.
62
1-8
1993
Fasciola hepatica
Cordova, M.; Jara, J.; Del Nery, E.; Hirata, I.Y.; Araujo, M.S.; Carmona, A.K.; Juliano, M.A.; Juliano, L.
Characterization of two cysteine proteinases secreted by Fasciola hepatica and demonstration of their kininogenase activity
Mol. Biochem. Parasitol.
116
109-115
2001
Fasciola hepatica
Dalton, J.P.; Neill, S.O.; Stack, C.; Collins, P.; Walshe, A.; et al.
Fasciola hepatica cathepsin L-like protease: biology, function, and potential in the development of the first generation liver fluke vaccines
Int. J. Parasitol.
33
1173-1181
2003
Fasciola hepatica
Kuk, S.; Kaplan, M.; Ozdarendeli, A.; Tonbak, S.; Felek, S.; Kalkan, A.
Fasciola hepatica cathepsin L1 from a Turkish isolate is related to Asiatic isolates
Acta Parasitol.
50
244-248
2005
Fasciola hepatica (Q2HPD3), Fasciola hepatica (Q7KYH5), Fasciola gigantica (Q8MUT6), Fasciola gigantica (Q9XYL8)
-
Ruth, D.M.; McMahon, G.; O'Fagain, C.
Peptide synthesis by recombinant Fasciola hepatica cathepsin L1
Biochimie
88
117-120
2006
Fasciola hepatica
Collins, P.R.; Stack, C.M.; ONeill, S.M.; Doyle, S.; Ryan, T.; Brennan, G.P.; Mousley, A.; Stewart, M.; Maule, A.G.; Dalton, J.P.; Donnelly, S.
Cathepsin L1, the major protease involved in liver fluke (Fasciola hepatica) virulence: propetide cleavage sites and autoactivation of the zymogen secreted from gastrodermal cells
J. Biol. Chem.
279
17038-17046
2004
Fasciola hepatica
Stack, C.M.; Donnelly, S.; Lowther, J.; Xu, W.; Collins, P.R.; Brinen, L.S.; Dalton, J.P.
The major secreted cathepsin L1 protease of the liver fluke, Fasciola hepatica: a Leu-12 to Pro-12 replacement in the nonconserved C-terminal region of the prosegment prevents complete enzyme autoactivation and allows definition of the molecular events in
J. Biol. Chem.
282
16532-16543
2007
Fasciola hepatica
Stack, C.M.; Caffrey, C.R.; Donnelly, S.M.; Seshaadri, A.; Lowther, J.; Tort, J.F.; Collins, P.R.; Robinson, M.W.; Xu, W.; McKerrow, J.H.; Craik, C.S.; Geiger, S.R.; Marion, R.; Brinen, L.S.; Dalton, J.P.
Structural and functional relationships in the virulence-associated cathepsin L proteases of the parasitic liver fluke, Fasciola hepatica
J. Biol. Chem.
283
9896-9908
2008
Fasciola hepatica (Q24940), Fasciola hepatica
Kesik, M.; Jedlina-Panasiuk, L.; Kozak-Cieszczyk, M.; P?ucienniczak, A.; Wedrychowicz, H.
Enteral vaccination of rats against Fasciola hepatica using recombinant cysteine proteinase (cathepsin L1)
Vaccine
25
3619-3628
2007
Fasciola hepatica
Berasain, P.; Carmona, C.; Frangione, B.; Dalton, J.P.; Goni, F.
Fasciola hepatica: parasite-secreted proteinases degrade all human IgG subclasses: determination of the specific cleavage sites and identification of the immunoglobulin fragments produced
Exp. Parasitol.
94
99-110
2000
Fasciola hepatica
Lowther, J.; Robinson, M.W.; Donnelly, S.M.; Xu, W.; Stack, C.M.; Matthews, J.M.; Dalton, J.P.
The importance of pH in regulating the function of the Fasciola hepatica cathepsin L1 cysteine protease
PLoS Negl. Trop. Dis.
3
e369
2009
Fasciola hepatica
Robinson, M.W.; Dalton, J.P.; Donnelly, S.
Helminth pathogen cathepsin proteases
Trends Biochem. Sci.
33
601-608
2008
Fasciola hepatica (Q24940)
Jayaraj, R.; Piedrafita, D.; Dynon, K.; Grams, R.; Spithill, T.W.; Smooker, P.M.
Vaccination against fasciolosis by a multivalent vaccine of stage-specific antigens
Vet. Parasitol.
160
230-236
2009
Fasciola hepatica
Robinson, M.W.; Tort, J.F.; Lowther, J.; Donnelly, S.M.; Wong, E.; Xu, W.; Stack, C.M.; Padula, M.; Herbert, B.; Dalton, J.P.
Proteomics and phylogenetic analysis of the cathepsin L protease family of the helminth pathogen Fasciola hepatica: expansion of a repertoire of virulence-associated factors
Mol. Cell. Proteomics
7
1111-1123
2008
Fasciola hepatica (Q7JNQ9)
Villa-Mancera, A.; Quiroz-Romero, H.; Correa, D.; Ibarra, F.; Reyes-Perez, M.; Reyes-Vivas, H.; Lopez-Velazquez, G.; Gazarian, K.; Gazarian, T.; Alonso, R.A.
Induction of immunity in sheep to Fasciola hepatica with mimotopes of cathepsin L selected from a phage display library
Parasitology
135
1437-1445
2008
Fasciola hepatica
Moran, B.W.; Anderson, F.P.; Ruth, D.M.; Fagain, C.O.; Dalton, J.P.; Kenny, P.T.
Fluorobenzoyl dipeptidyl derivatives as inhibitors of the Fasciola hepatica cysteine protease cathepsin L1
J. Enzyme Inhib. Med. Chem.
25
1-12
2010
Fasciola hepatica
Golden, O.; Flynn, R.J.; Read, C.; Sekiya, M.; Donnelly, S.M.; Stack, C.; Dalton, J.P.; Mulcahy, G.
Protection of cattle against a natural infection of Fasciola hepatica by vaccination with recombinant cathepsin L1 (rFhCL1)
Vaccine
28
5551-5557
2010
Fasciola hepatica
Perez-Ecija, R.A.; Mendes, R.E.; Zafra, R.; Buffonni, L.; Martinez-Moreno, A.; Perez, J.
Pathological and parasitological protection in goats immunised with recombinant cathepsin L1 and challenged with Fasciola hepatica
Vet. J.
185
351-353
2010
Fasciola hepatica
Buffoni, L.; Martinez-Moreno, F.J.; Zafra, R.; Mendes, R.E.; Perez-Ecija, A.; Sekiya, M.; Mulcahy, G.; Perez, J.; Martinez-Moreno, A.
Humoral immune response in goats immunised with cathepsin L1, peroxiredoxin and Sm14 antigen and experimentally challenged with Fasciola hepatica
Vet. Parasitol.
185
315-321
2012
Fasciola hepatica
Hernandez-Guzman, K.; Sahagun-Ruiz, A.; Vallecillo, A.J.; Cruz-Mendoza, I.; Quiroz-Romero, H.
Construction and evaluation of a chimeric protein made from Fasciola hepatica leucine aminopeptidase and cathepsin L1
J. Helminthol.
90
7-13
2016
Fasciola hepatica (Q9GRW5), Fasciola hepatica
Meshgi, B.; Jalousian, F.; Fathi, S.; Jahani, Z.
Design and synthesis of a new peptide derived from Fasciola gigantica cathepsin L1 with potential application in serodiagnosis of fascioliasis
Exp. Parasitol.
189
76-86
2018
Fasciola gigantica (Q9XYL8), Fasciola gigantica, Fasciola hepatica (Q24940)
Hernandez-Guzman, K.; Sahagun-Ruiz, A.; Vallecillo, A.; Cruz-Mendoza, I.; Quiroz-Romero, H.
Construction and evaluation of a chimeric protein made from Fasciola hepatica leucine aminopeptidase and cathepsin L1
J. Helminthol.
90
7-13
2016
Fasciola hepatica (Q9GRW5), Fasciola hepatica
Mebius, M.M.; Op Heij, J.M.J.; Tielens, A.G.M.; de Groot, P.G.; Urbanus, R.T.; van Hellemond, J.J.
Fibrinogen and fibrin are novel substrates for Fasciola hepatica cathepsin L peptidases
Mol. Biochem. Parasitol.
221
10-13
2018
Fasciola hepatica (H8WG11), Fasciola hepatica
Martinez-Sernandez, V.; Perteguer, M.J.; Hernandez-Gonzalez, A.; Mezo, M.; Gonzalez-Warleta, M.; Orbegozo-Medina, R.A.; Romaris, F.; Paniagua, E.; Garate, T.; Ubeira, F.M.
Comparison of recombinant cathepsins L1, L2, and L5 as ELISA targets for serodiagnosis of bovine and ovine fascioliasis
Parasitol. Res.
117
1521-1534
2018
Fasciola hepatica (H8WG11)
Ortega-Vargas, S.; Espitia, C.; Sahagun-Ruiz, A.; Parada, C.; Balderas-Loaeza, A.; Villa-Mancera, A.; Quiroz-Romero, H.
Moderate protection is induced by a chimeric protein composed of leucine aminopeptidase and cathepsin L1 against Fasciola hepatica challenge in sheep
Vaccine
37
3234-3240
2019
Fasciola hepatica (Q9GRW5), Fasciola hepatica
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