Information on EC 3.4.22.B62 - cathepsin L3

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The expected taxonomic range for this enzyme is: Fasciola hepatica

EC NUMBER
COMMENTARY hide
3.4.22.B62
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
cathepsin L3
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
70fold preference for tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin over typical cathepsin substrates with hydrophobic or aliphatic residues in the S2 position. Efficiently cleaves type I collagen over different pH and temperature conditions
show the reaction diagram
-
-
-
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-L-Phe-L-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-L-Val-L-Leu-L-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-Val-Leu-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
Boc-AGPR-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
Boc-VLK-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
Boc-VPR-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
Collagen + H2O
?
show the reaction diagram
human immumglobulin G + H2O
?
show the reaction diagram
partial cleavage
-
-
?
tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
restricted substrate specificity with 70fold preference for tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin over benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin and benzyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin
-
-
?
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin + H2O
tosyl-Gly-Pro-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
tosyl-GPK-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
tosyl-GPR-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
Type I collagen + H2O
?
show the reaction diagram
type II collagen + H2O
?
show the reaction diagram
-
cathepsin L3 exhibits collagenase activity by cleaving at multiple sites within the alpha1 and alpha2 triple helix regions (Col domains)
-
-
?
Z-Gly-Pro-Gly-Gly-Pro-Ala + H2O
Z-Gly-Pro + Gly + Gly + L-Pro-L-Ala
show the reaction diagram
-
-
-
-
?
Z-Gly-Pro-Leu-Gly-Pro + H2O
Z-Gly-Pro + L-Leu + Gly + L-Pro
show the reaction diagram
-
-
-
-
?
Z-Leu-Arg-NHMe + H2O
?
show the reaction diagram
-
-
-
-
?
Z-Phe-Arg-NHMe + H2O
?
show the reaction diagram
-
-
-
-
?
Z-Val-Leu-Lys-7-amido-4-methylcoumarin
Z-Val-Leu-Lys + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
-
Z-VVR-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
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 + H2O
?
show the reaction diagram
B9VXS1, B9VXS2
-
-
-
?
human immumglobulin G + H2O
?
show the reaction diagram
B9VXS1, B9VXS2
partial cleavage
-
-
?
Type I collagen + H2O
?
show the reaction diagram
additional information
?
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cathepsin K inhibitor II
-
i.e. Z-LNHNHCONHNHLF-Boc
-
Z-Phe-Ala-CHN2
-
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.02353
benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin
-
in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C
0.0157
benzyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin
-
in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C
0.0235
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
-
-
0.0157
benzyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin
-
-
0.0096
Boc-AGPR-7-amido-4-methylcoumarin
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
0.00613
Boc-VLK-7-amido-4-methylcoumarin
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
0.0211
Boc-VPR-7-amido-4-methylcoumarin
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
0.0041
tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin
-
in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C
0.0041 - 0.0067
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
0.0098
tosyl-GPK-7-amido-4-methylcoumarin
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
0.0106
tosyl-GPR-7-amido-4-methylcoumarin
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
0.048
Z-Leu-Arg-NHMe
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
0.02005
Z-Phe-Arg-NHMe
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
0.0157 - 0.0448
Z-Val-Leu-Lys-7-amido-4-methylcoumarin
0.0117
Z-VVR-7-amido-4-methylcoumarin
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.39
benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C
0.33
benzyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C
0.39
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin
Fasciola hepatica
-
-
0.33
benzyloxycarbonyl-Val-Leu-Lys-7-amido-4-methylcoumarin
Fasciola hepatica
-
-
0.58
Boc-AGPR-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
0.01
Boc-VLK-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
0.033
Boc-VPR-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
6
tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C
0.0053 - 6
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
0.15
tosyl-GPK-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
1.015
tosyl-GPR-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
1.2
Z-Leu-Arg-NHMe
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
0.02
Z-Phe-Arg-NHMe
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
0.03 - 0.33
Z-Val-Leu-Lys-7-amido-4-methylcoumarin
0.043
Z-VVR-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
16.78
benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C
11711
20.76
benzyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C
41331
60.76
Boc-AGPR-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
19614
1.748
Boc-VLK-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
19609
1.564
Boc-VPR-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
19613
1464
tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 50 mM sodium phosphate, pH 6.5, 1 mM dithiothreitol, 1 mM EDTA, at 24°C
8882
1.017 - 1464
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin
2114
36.42
tosyl-GPK-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
19612
95.77
tosyl-GPR-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
19611
25
Z-Leu-Arg-NHMe
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
19607
0.9993
Z-Phe-Arg-NHMe
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
19608
0.59 - 20.76
Z-Val-Leu-Lys-7-amido-4-methylcoumarin
202466
3.697
Z-VVR-7-amido-4-methylcoumarin
Fasciola hepatica
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
19610
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00000905
cathepsin K inhibitor II
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
-
0.0003365
Z-Phe-Ala-CHN2
-
in 100 mM sodium phosphate buffer (pH 6.0) containing 1 mM dithiothreitol and 1 mM EDTA, at 37°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6
-
using benzyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin as substrate
7
-
; using tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin as substrate
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 9
-
CL3 has about 70% of the maximum activity at pH 5.0 and about 40% of the maximum activity at pH 9.0 using benzyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin as substrate
5.5 - 8
-
CL3 has 35% of the maximum activity at pH 5.5 and about 70% of the maximum activity at pH 8.0 using tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin as substrate
6 - 8
-
pH 6: about 70% of maximal activity, pH 8.0: abput 70% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.18
-
pro-cathepsin L3, calculated from sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
24500
-
x * 24500, mature enzyme, SDS-PAGE
35550
-
pro-cathepsin L3, calculated from sequence
38080
x * 38080, isoform CL3-1, calculated from amino acid sequence
38280
x * 38280, isoform CL3-2, calculated from amino acid sequence
46000
x * 46000, recombinant proenzyme, SDS-PAGE; x * 46000, recombinant proenzyme, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
the propeptide of CL3 plays an important role in inducing immunity against Fasciola hepatica infection; the propeptide of CL3 plays an important role in inducing immunity against Fasciola hepatica infection
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
-
37°C, 50% loss of activity after 8 h, 64% loss of activity after 24 h; 50% of the activity is conserved after 8 h incubation and 36% of it is retained after 24 h incubation at pH 7.0 and 37°C
694073
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; HiTrap QXL column chromatography, gel filtration
-
Ni-NTA-agarose column chromatography
-
nickel affinity column chromatography; nickel affinity column chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
baculovirus produced procathepsin L3 partly protects rats against fasciolosis, whereas the same coding region expressed in yeast does not
-
expressed in Escherichia coli BL21 and Pichia pastoris X33 cells; expressed in Escherichia coli BL21 and Pichia pastoris X33 cells
expressed in Hansenula polymorpha
-
expressed in Hansenula polymorpha strain NCYC 495; the proenzyme is expressed in the methylotrophic yeast Hansenula polymorpha and the mature enzyme is obtained from the culture medium
-
expressed in Pichia angusta
-
recombinant expression of a CL3 cDNA in Saccharomyces cerevisiae results in secretion of the proenzyme form; recombinant expression of a CL3 cDNA in Saccharomyces cerevisiae results in secretion of the proenzyme form
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
maximum expression level is observed in newly excysted juveniles immediately after excystment; maximum expression level is observed in newly excysted juveniles immediately after excystment
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H61N/W67L
-
the double mutant presents a profile very similar to the W67L single mutant
W67L
-
the mutation renders the enzyme incapable of digesting collagen. The mutant shows drastic diminution in specificity towards tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin (1440fold) and also large (35fold) decrease in specificity towards Z-Val-Leu-Lys-7-amido-4-methylcoumarin