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Information on EC 3.4.22.B61 - cathepsin L5
for references in articles please use BRENDA:EC3.4.22.B61preliminary BRENDA-supplied EC number
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EC Tree
3.4.22.B61 cathepsin L5Specify your search results
Word Map
- 3.4.22.B61
- fasciola
- cathepsins
- hepatica
- fluke
-
subsite
- medicine
-
evasion
-
l-like
- gigantica
-
helminth
- cattle
- sheep
-
metacercariae
-
stage-specific
-
excysted
-
multivalent
-
veterinary
- agriculture
- diagnostics
The expected taxonomic range for this enzyme is: Fasciola
Reaction Schemes
cathepsin L5 does not exhibit substantial catalytic activity against substrates containing proline in the P2 position (thereby distinguished from Fasciola cathepsin L1, L2 and L3)
Synonyms
fhcatl5, cathepsin l5, fhcl5, cat l5, 
more
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
cathepsin L5
FhCatL5
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cathepsin L5 does not exhibit substantial catalytic activity against substrates containing proline in the P2 position (thereby distinguished from Fasciola cathepsin L1, L2 and L3)
-
-
-
-
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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-Phe-L-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-L-Phe-L-Arg + 7-amino-4-methylcoumarin
CatL5 has a clear preference for benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin over tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin
-
-
?
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
-
-
-
?
Hemoglobin + H2O
?
cathepsin l5 hydrolyzes hemoglobin only at pH 5.5
-
-
?
tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin + H2O
tosyl-Gly-L-Pro-L-Arg + 7-amino-4-methylcoumarin
weak substrate
-
-
?
tosyl-Gly-Phe-Arg-7-amido-4-methylcoumarin + H2O
tosyl-Gly-Phe-Arg + 7-amino-4-methylcoumarin
-
-
-
?
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin + H2O
tosyl-Gly-Pro-Arg + 7-amino-4-methylcoumarin
-
-
-
?
type 1 collagen + H2O
?
efficient cleavage of type 1 collagen by CatL5 is limited to the beta-and gamma-chains
-
-
?
Z-Phe-Arg-7-amido-4-methylcoumarin + H2O
Z-Phe-Arg + 7-amino-4-methylcoumarin
preferred substrate
-
-
?
additional information
?
-
cathepsin L5 does not exhibit substantial catalytic activity against substrates containing proline in the P2 position
-
-
?
additional information
?
-
-
cathepsin L5 does not exhibit substantial catalytic activity against substrates containing proline in the P2 position
-
-
?
additional information
?
-
no activity with PFR-7-amido-4-methylcoumarin, Boc-DPR-7-amido-4-methylcoumarin, Boc-AGPR-7-amido-4-methylcoumarin, Boc-VPR-7-amido-4-methylcoumarin, D-Val-Leu-Arg-7-amido-4-methylcoumarin, Boc-VLK-7-amido-4-methylcoumarin, and benzoyl-FVR-7-amido-4-methylcoumarin
-
-
?
additional information
?
-
-
no activity with PFR-7-amido-4-methylcoumarin, Boc-DPR-7-amido-4-methylcoumarin, Boc-AGPR-7-amido-4-methylcoumarin, Boc-VPR-7-amido-4-methylcoumarin, D-Val-Leu-Arg-7-amido-4-methylcoumarin, Boc-VLK-7-amido-4-methylcoumarin, and benzoyl-FVR-7-amido-4-methylcoumarin
-
-
?
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.02481
benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin
wild type enzyme, at pH 5.5 and 37°C
0.1373
benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin
mutant enzyme L69W, at pH 5.5 and 37°C
0.0059
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
FhCatL5-L69Y
0.05392
tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin
wild type enzyme, at pH 5.5 and 37°C
0.0786
tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin
mutant enzyme L69W, at pH 5.5 and 37°C
0.05193
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
mutant enzyme L69Y, at pH 5.5 and 37°C
0.06346
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
mutant enzyme L69Y, at pH 5.5 and 37°C
0.07674
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
mutant enzyme L69C, at pH 5.5 and 37°C
0.1269
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
mutant enzyme L69S, at pH 5.5 and 37°C
5392
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
wild type enzyme, at pH 5.5 and 37°C
0.01272
Z-Phe-Arg-7-amido-4-methylcoumarin
mutant enzyme L69S, at pH 5.5 and 37°C
0.01372
Z-Phe-Arg-7-amido-4-methylcoumarin
mutant enzyme L69C, at pH 5.5 and 37°C
0.01911
Z-Phe-Arg-7-amido-4-methylcoumarin
mutant enzyme L69F, at pH 5.5 and 37°C
0.02481
Z-Phe-Arg-7-amido-4-methylcoumarin
wild type enzyme, at pH 5.5 and 37°C
0.03287
Z-Phe-Arg-7-amido-4-methylcoumarin
mutant enzyme L69Y, at pH 5.5 and 37°C
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
11.05
benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin
mutant enzyme L69W, at pH 5.5 and 37°C
17.89
benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin
wild type enzyme, at pH 5.5 and 37°C
2145
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
FhCatL5-L69Y
0.84
tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin
wild type enzyme, at pH 5.5 and 37°C
1.34
tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin
mutant enzyme L69W, at pH 5.5 and 37°C
0.19
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
mutant enzyme L69C, at pH 5.5 and 37°C
0.27
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
mutant enzyme L69S, at pH 5.5 and 37°C
0.84
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
wild type enzyme, at pH 5.5 and 37°C
1.78
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
mutant enzyme L69Y, at pH 5.5 and 37°C
2.54
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
mutant enzyme L69Y, at pH 5.5 and 37°C
1.73
Z-Phe-Arg-7-amido-4-methylcoumarin
mutant enzyme L69S, at pH 5.5 and 37°C
1.92
Z-Phe-Arg-7-amido-4-methylcoumarin
mutant enzyme L69C, at pH 5.5 and 37°C
17.89
Z-Phe-Arg-7-amido-4-methylcoumarin
wild type enzyme, at pH 5.5 and 37°C
17.93
Z-Phe-Arg-7-amido-4-methylcoumarin
mutant enzyme L69F, at pH 5.5 and 37°C
21.52
Z-Phe-Arg-7-amido-4-methylcoumarin
mutant enzyme L69Y, at pH 5.5 and 37°C
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
80.5
benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin
mutant enzyme L69W, at pH 5.5 and 37°C
721
benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin
wild type enzyme, at pH 5.5 and 37°C
15.5
tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin
wild type enzyme, at pH 5.5 and 37°C
17
tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin
mutant enzyme L69W, at pH 5.5 and 37°C
2.1
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
mutant enzyme L69S, at pH 5.5 and 37°C
2.5
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
mutant enzyme L69C, at pH 5.5 and 37°C
15.5
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
wild type enzyme, at pH 5.5 and 37°C
34.4
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
mutant enzyme L69Y, at pH 5.5 and 37°C
40
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
mutant enzyme L69Y, at pH 5.5 and 37°C
136
Z-Phe-Arg-7-amido-4-methylcoumarin
mutant enzyme L69S, at pH 5.5 and 37°C
140
Z-Phe-Arg-7-amido-4-methylcoumarin
mutant enzyme L69C, at pH 5.5 and 37°C
655
Z-Phe-Arg-7-amido-4-methylcoumarin
mutant enzyme L69Y, at pH 5.5 and 37°C
721
Z-Phe-Arg-7-amido-4-methylcoumarin
wild type enzyme, at pH 5.5 and 37°C
938
Z-Phe-Arg-7-amido-4-methylcoumarin
mutant enzyme L69F, at pH 5.5 and 37°C
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
additional information
additional information
cathepsin L5 (FhCL5) constitutes approximately 7% and 5-10% of the secreted protease content of immature and adult flukes, respectively
brenda
additional information
-
cathepsin L5 (FhCL5) constitutes approximately 7% and 5-10% of the secreted protease content of immature and adult flukes, respectively
brenda
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
the enzyme is secreted. Cathepsin L5 (FhCL5) constitutes approximately 7% and 5-10% of the secreted protease content of immature and adult flukes, respectively
-
brenda
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
adult flukes produce three different clades of cathepsin Ls: FhCL1, FhCL2, and FhCL5
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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). Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
L69F
the mutant shows a 2-3fold increase in the ability to cleave substrates with P2 proline
L69W
the mutation results in an increased preference for substrates containing proline at P2. The mutant shows increased Km values for benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin and tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin compared to the wild type enzyme, but show increased catalytic efficiency for tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin
L69Y
L69Y
substantial increase in the ability to accommodate substrates with a proline residue in the P2
L69Y
the mutant shows a 2-3fold increase in the ability to cleave substrates with P2 proline
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.5
wild type CatL5 and mutant CatL5 L69W retain more than 90% activity after 4 h incubation at pH 7.5 and about 40% activity after incubation for 24 h
717293
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
nickel chelating column chromatography and Mono S column chromatography
nickel-chelating Sepharose column chromatography and Mono S column chromatography
recombinant FLAG- and His6-tagged enzyme from Saccharomyces cerevisiae strain BJ3505 by nickel affinity chromatography
recombinant solubilized and refolded His-tagged enzyme from Escherichia coli by nickel affinity chromatography
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
cathepsin L5 is expressed in COS 7 cells, it induces a humoral response when delivered as secretory constructs in mice
-
gene FhpCL5, DNA and amino acid sequence determination and analysis, sequence comparisons of rFhpCLs, recombinant expression of His-tagged enzyme in Escherichia coli
recombinant expression of FLAG- and His6-tagged enzyme in Saccharomyces cerevisiae strain BJ3505
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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
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
-
vaccination of rats with recombinant zymogen and subsequent challenge with Fasciola hepatica metercercariae. Vaccination results in significantly smaller and fewer flukes than in controls. Maximal protection of 83% is seen in the group vaccinated with cathepsin B and cathepsin L5 in combinantion
agriculture
Fasciola hepatica infection continues to be a major problem in the agriculture sector, particularly in sheep and cattle. Cathepsin L and B proteases are major components of the excretory/secretory material of the parasite, and their roles in several important aspects of parasite invasion and survival has led to their use as targets in rational vaccine design. Recombinant versions of cathepsin L5 and cathepsin B2 produced in yeast are used in combination in a commercial adjuvant preparation to vaccinate sheep. Intramuscular and intranasal forms of administration are applied, and sheep are subsequently challenged with 150 Fasciola hepatica metacercariae. Intramuscular vaccination is able to induce a strong systemic antibody response against both antigens, but fails to confer significant protection. Conversely, no elevated antibody response is detected against the vaccine antigens following nasal vaccination, but a reduction in parasite egg viability (above 92%) and a statistically significant, predominantly adjuvant-mediated reduction in worm burdens is observed
medicine
-
cathepsin L5 antibody titre correlates well with actual Fasciola worm burdens
medicine
-
cathepsin L5, cathepsin L1g and cathepsin B are expressed in yeast from cDNA clones isolated from adult, metacercariae and newly excysted juvenile flukes respectively. Each is used solely or in combination to vaccinate rats that are subsequently challenged with Fasciola hepatica metercercariae. Each protein induces an immune response, and all groups vaccinated with recombinant protein yield significantly fewer and smaller flukes than the control group. Maximal protection of 83% is seen in the group vaccinated with cathepsin B and cathepsin L5 in combination
medicine
Fasciola hepatica infection continues to be a major problem in the agriculture sector, particularly in sheep and cattle. Cathepsin L and B proteases are major components of the excretory/secretory material of the parasite, and their roles in several important aspects of parasite invasion and survival has led to their use as targets in rational vaccine design. Recombinant versions of cathepsin L5 and cathepsin B2 produced in yeast are used in combination in a commercial adjuvant preparation to vaccinate sheep. Intramuscular and intranasal forms of administration are applied, and sheep are subsequently challenged with 150 Fasciola hepatica metacercariae. Intramuscular vaccination is able to induce a strong systemic antibody response against both antigens, but fails to confer significant protection. Conversely, no elevated antibody response is detected against the vaccine antigens following nasal vaccination, but a reduction in parasite egg viability (above 92%) and a statistically significant, predominantly adjuvant-mediated reduction in worm burdens is observed
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Smooker, P.M.; Kennedy, N.J.; Steeper, K.R.; Christopoulos, H.; Spithill, T.W.
Fasciola: kinetics and quality of humoral responses to fatty acid binding protein and cathepsin L following delivery as DNA vaccines in mice
Exp. Parasitol.
97
154-160
2001
Fasciola hepatica
brenda
Smooker, P.M.; Whisstock, J.C.; Irving, J.A.
Siyaguna, S.; Spithill, T.W.: Pike, R.N.: A single amino acid substitution affects substrate specificity in cysteine proteinases from Fasciola hepatica
Protein Sci.
9
2567-2572
2000
Fasciola hepatica (Q9NB30), Fasciola hepatica
brenda
Raadsma, H.W.; Kingsford, N.M.; Suharyanta; Spithill, T.W.; Piedrafita, D.
Host responses during experimental infection with Fasciola gigantica and Fasciola hepatica in Merino sheep II. Development of a predictive index for Fasciola gigantica worm burden
Vet. Parasitol.
154
250-261
2008
Fasciola gigantica
brenda
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
brenda
Norbury, L.J.; Beckham, S.; Pike, R.N.; Grams, R.; Spithill, T.W.; Fecondo, J.V.; Smooker, P.M.
Adult and juvenile Fasciola cathepsin L proteases: different enzymes for different roles
Biochimie
93
604-611
2011
Fasciola hepatica (Q9NB30), Fasciola hepatica
brenda
Norbury, L.J.; Hung, A.; Beckham, S.; Pike, R.N.; Spithill, T.W.; Craik, C.S.; Choe, Y.; Fecondo, J.V.; Smooker, P.M.
Analysis of Fasciola cathepsin L5 by S2 subsite substitutions and determination of the P1-P4 specificity reveals an unusual preference
Biochimie
94
1119-1127
2012
Fasciola hepatica (Q9NB30)
brenda
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 (A0A2H4PKA2), Fasciola hepatica
brenda
Norbury, L.J.; Basalaj, K.; Zawistowska-Deniziak, A.; Sielicka, A.; Wilkowski, P.; Wesolowska, A.; Smooker, P.M.; Wedrychowicz, H.
Intranasal delivery of a formulation containing stage-specific recombinant proteins of Fasciola hepatica cathepsin L5 and cathepsin B2 triggers an anti-fecundity effect and an adjuvant-mediated reduction in fluke burden in sheep
Vet. Parasitol.
258
14-23
2018
Fasciola hepatica (A0A2H4PKA2), Fasciola hepatica
brenda
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