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Literature summary for 3.4.22.15 extracted from

  • 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 (2011), Biochimie, 93, 604-611.
    View publication on PubMed
Search for more literature for 3.4.22.15

Activating Compound

Activating Compound Comment Organism Structure
cysteine activation with 10 mM cysteine-HCl for 2 h at 37°C Fasciola hepatica
cysteine activation with 10 mM cysteine-HCl for 2 h at 37°C Fasciola gigantica

Cloned(Commentary)

Cloned (Comment) Organism
expressed in yeast Fasciola hepatica
expressed in yeast Fasciola gigantica

Protein Variants

Protein Variants Comment Organism
L69W isoform CatL5 mutant, compared to the wild type enzyme the mutant has a higher efficiency of cleavage (kcat) against tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin Fasciola hepatica

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
Albumin + H2O Fasciola hepatica isoform CatL5 shows weak activity against albumin ?
-
 ?
Fibronectin + H2O Fasciola hepatica cleavage occurs only at pH 5.5 ?
-
 ?
Fibronectin + H2O Fasciola gigantica cleavage occurs only at pH 5.5 ?
-
 ?
Immunoglobulin + H2O Fasciola hepatica
-
 ?
-
 ?
Immunoglobulin + H2O Fasciola gigantica
-
 ?
-
 ?
Laminin + H2O Fasciola hepatica cleavage occurs only at pH 5.5 ?
-
 ?
Laminin + H2O Fasciola gigantica cleavage occurs only at pH 5.5 ?
-
 ?
additional information Fasciola hepatica isoform CatL1G can cleave substrates with proline residues at P2 ?
-
 ?
additional information Fasciola gigantica isoform CatL1G can cleave substrates with proline residues at P2, isoform CatL1G is unable to digest albumin or hemoglobin, efficiently at either pH 5.5 or 7.0 ?
-
 ?
type 1 collagen + H2O Fasciola hepatica efficient cleavage of type 1 collagen by isoform CatL5 is limited to the beta-and gamma-chains ?
-
 ?
type 1 collagen + H2O Fasciola gigantica isoform Catl1G completely digests collagen type 1 ?
-
 ?

Organism

Organism UniProt Comment Textmining
Fasciola gigantica
-
-
-
Fasciola hepatica
-
-
-

Purification (Commentary)

Purification (Comment) Organism
nickel-chelating Sepharose column chromatography Fasciola gigantica
nickel-chelating Sepharose column chromatography, gel filtration Fasciola hepatica

Source Tissue

Source Tissue Comment Organism Textmining

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Albumin + H2O isoform CatL5 shows weak activity against albumin Fasciola hepatica ?
-
 ?
benzoyl-L-Phe-L-Val-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica benzoyl-L-Phe-L-Val-L-Arg + 7-amino-4-methylcoumarin
-
 ?
benzoyl-L-Phe-L-Val-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola gigantica benzoyl-L-Phe-L-Val-L-Arg + 7-amino-4-methylcoumarin
-
 ?
D-Val-Leu-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica D-Val-Leu-Arg + 7-amino-4-methylcoumarin
-
 ?
D-Val-Leu-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola gigantica D-Val-Leu-Arg + 7-amino-4-methylcoumarin
-
 ?
Fibronectin + H2O cleavage occurs only at pH 5.5 Fasciola hepatica ?
-
 ?
Fibronectin + H2O cleavage occurs only at pH 5.5 Fasciola gigantica ?
-
 ?
Immunoglobulin + H2O
-
 Fasciola hepatica ?
-
 ?
Immunoglobulin + H2O
-
 Fasciola gigantica ?
-
 ?
L-Phe-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola gigantica L-Phe-L-Arg + 7-amino-4-methylcoumarin
-
 ?
L-Phe-L-Arg-7-amido-4-methylcoumarin + H2O preferred substrate for isoform CatL5 Fasciola hepatica L-Phe-L-Arg + 7-amino-4-methylcoumarin
-
 ?
L-Pro-L-Phe-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica L-Pro-L-Phe-L-Arg + 7-amino-4-methylcoumarin
-
 ?
L-Pro-L-Phe-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola gigantica L-Pro-L-Phe-L-Arg + 7-amino-4-methylcoumarin
-
 ?
Laminin + H2O cleavage occurs only at pH 5.5 Fasciola hepatica ?
-
 ?
Laminin + H2O cleavage occurs only at pH 5.5 Fasciola gigantica ?
-
 ?
additional information isoform CatL1G can cleave substrates with proline residues at P2 Fasciola hepatica ?
-
 ?
additional information isoform CatL1G can cleave substrates with proline residues at P2, isoform CatL1G is unable to digest albumin or hemoglobin, efficiently at either pH 5.5 or 7.0 Fasciola gigantica ?
-
 ?
tert-butyloxycarbonyl-L-Ala-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica tert-butyloxycarbonyl-L-Ala-Gly-L-Pro-L-Arg + 7-amino-4-methylcoumarin
-
 ?
tert-butyloxycarbonyl-L-Ala-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola gigantica tert-butyloxycarbonyl-L-Ala-Gly-L-Pro-L-Arg + 7-amino-4-methylcoumarin
-
 ?
tert-butyloxycarbonyl-L-Asp-L-Pro-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica tert-butyloxycarbonyl-L-Asp-L-Pro-L-Arg + 7-amino-4-methylcoumarin
-
 ?
tert-butyloxycarbonyl-L-Asp-L-Pro-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola gigantica tert-butyloxycarbonyl-L-Asp-L-Pro-L-Arg + 7-amino-4-methylcoumarin
-
 ?
tert-butyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica tert-butyloxycarbonyl-L-Val-L-Leu-L-Lys + 7-amino-4-methylcoumarin
-
 ?
tert-butyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin + H2O
-
 Fasciola gigantica tert-butyloxycarbonyl-L-Val-L-Leu-L-Lys + 7-amino-4-methylcoumarin
-
 ?
tert-butyloxycarbonyl-L-Val-L-Pro-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica tert-butyloxycarbonyl-L-Val-L-Pro-L-Arg + 7-amino-4-methylcoumarin
-
 ?
tert-butyloxycarbonyl-L-Val-L-Pro-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola gigantica tert-butyloxycarbonyl-L-Val-L-Pro-L-Arg + 7-amino-4-methylcoumarin
-
 ?
tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica tosyl-Gly-L-Pro-L-Arg + 7-amino-4-methylcoumarin
-
 ?
tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin + H2O preferred substrate for isoform CatL1G Fasciola gigantica tosyl-Gly-L-Pro-L-Arg + 7-amino-4-methylcoumarin
-
 ?
type 1 collagen + H2O efficient cleavage of type 1 collagen by isoform CatL5 is limited to the beta-and gamma-chains Fasciola hepatica ?
-
 ?
type 1 collagen + H2O isoform Catl1G completely digests collagen type 1 Fasciola gigantica ?
-
 ?

Subunits

Subunits Comment Organism
? x * about 38000, pro-enzyme, SDS-PAGE Fasciola hepatica
? x * about 38000, pro-enzyme, SDS-PAGE Fasciola gigantica

Synonyms

Synonyms Comment Organism
CatL1G juvenile isoform Fasciola gigantica
CatL5 adult isoform Fasciola hepatica

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6 7
-
 Fasciola hepatica
6 7
-
 Fasciola gigantica

pH Stability

pH Stability pH Stability Maximum Comment Organism
4 9 after incubation for 2 h at 37°C in AMT buffers (100 mM sodium acetate, 100 mM MES, 200 mM Tris, 1 mM EDTA), the isoform CatL1G shows residual activities of about 30% at pH 4.0, about 75% at pH 5.0, about 85% at pH 6.0, about 45% at pH 7.0, and about 10% at pH 8.0-9.0. Isoform CatL1G retains 25% activity after 2 h at pH 7.5, but retains over 80% activity over 4 h at pH 5.5 and 50% over 24 h Fasciola gigantica
4 9 isoform CatL5 undergoes significant autoprocessing if the pH of the yeast media is allowed to drop below pH 6.0. After incubation for 2 h at 37°C in AMT buffers (100 mM sodium acetate, 100 mM MES, 200 mM Tris, 1 mM EDTA), the isoform CatL5 shows residual activities of about 30% at pH 4.0-5.0, about 60% at pH 6.0, about 85% at pH 7.0, about 100% at pH 8.0, and about 60% at pH 9.0. Isoform 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 Fasciola hepatica

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.01
-
 tert-butyloxycarbonyl-L-Val-L-Pro-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola gigantica
6.75
-
 tert-butyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola gigantica
10.1
-
 tert-butyloxycarbonyl-L-Asp-L-Pro-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
11.3
-
 tert-butyloxycarbonyl-L-Asp-L-Pro-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola gigantica
16.6
-
 tert-butyloxycarbonyl-L-Ala-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
18.1
-
 benzoyl-L-Phe-L-Val-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola gigantica
22.4
-
 tert-butyloxycarbonyl-L-Val-L-Pro-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
24.4
-
 tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
33.8
-
 tert-butyloxycarbonyl-L-Ala-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola gigantica
38.3
-
 D-Val-Leu-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola gigantica
38.5
-
 tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola gigantica
52.7
-
 tert-butyloxycarbonyl-L-Val-L-Pro-L-Arg-7-amido-4-methylcoumarin mutant enzyme L69W, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
75.2
-
 tert-butyloxycarbonyl-L-Asp-L-Pro-L-Arg-7-amido-4-methylcoumarin mutant enzyme L69W, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
84.3
-
 tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin mutant enzyme L69W, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
119
-
 tert-butyloxycarbonyl-L-Ala-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin mutant enzyme L69W, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
149
-
 L-Pro-L-Phe-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
152
-
 L-Phe-L-Arg-7-amido-4-methylcoumarin mutant enzyme L69W, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
182
-
 benzoyl-L-Phe-L-Val-L-Arg-7-amido-4-methylcoumarin mutant enzyme L69W, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
211
-
 L-Pro-L-Phe-L-Arg-7-amido-4-methylcoumarin mutant enzyme L69W, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
308
-
 D-Val-Leu-Arg-7-amido-4-methylcoumarin mutant enzyme L69W, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
474
-
 L-Phe-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
781
-
 benzoyl-L-Phe-L-Val-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
1390
-
 tert-butyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin mutant enzyme L69W, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
5420
-
 D-Val-Leu-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica
7220
-
 tert-butyloxycarbonyl-L-Val-L-Leu-L-Lys-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5, in the presence of 10 mM cysteine-HCl, at 37°C Fasciola hepatica