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Literature summary for 3.4.22.B62 extracted from

  • Corvo, I.; ODonoghue, A.J.; Pastro, L.; Pi-Denis, N.; Eroy-Reveles, A.; Roche, L.; McKerrow, J.H.; Dalton, J.P.; Craik, C.S.; Caffrey, C.R.; Tort, J.F.
    Dissecting the active site of the collagenolytic cathepsin L3 protease of the invasive stage of Fasciola hepatica (2013), PLoS Negl. Trop. Dis., 7, e2269.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.22.B62

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Hansenula polymorpha Fasciola hepatica

Protein Variants

Protein Variants Comment Organism
H61N/W67L the double mutant presents a profile very similar to the W67L single mutant Fasciola hepatica
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 Fasciola hepatica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0041
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin wild type enzyme, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica
0.0052
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme W67L, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica
0.0067
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme H61N/W67L, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica
0.0157
-
 Z-Val-Leu-Lys-7-amido-4-methylcoumarin wild type enzyme, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica
0.0287
-
 Z-Val-Leu-Lys-7-amido-4-methylcoumarin mutant enzyme H61N/W67L, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica
0.0448
-
 Z-Val-Leu-Lys-7-amido-4-methylcoumarin mutant enzyme W67L, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
Type I collagen + H2O Fasciola hepatica the enzyme with collagenolytic activity favours Pro at P2 and shows a distinctive specificity at the S3 pocket, accommodating preferentially the small Gly residue ?
-
 ?

Organism

Organism UniProt Comment Textmining
Fasciola hepatica
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica tosyl-Gly-Pro-Arg + 7-amino-4-methylcoumarin
-
 ?
Type I collagen + H2O the enzyme with collagenolytic activity favours Pro at P2 and shows a distinctive specificity at the S3 pocket, accommodating preferentially the small Gly residue Fasciola hepatica ?
-
 ?
Z-Val-Leu-Lys-7-amido-4-methylcoumarin
-
 Fasciola hepatica Z-Val-Leu-Lys + 7-amino-4-methylcoumarin
-
 ?

Synonyms

Synonyms Comment Organism
CL3
-
 Fasciola hepatica

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0053
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme W67L, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica
0.0072
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme H61N/W67L, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica
0.03
-
 Z-Val-Leu-Lys-7-amido-4-methylcoumarin mutant enzyme H61N/W67L, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica
0.03
-
 Z-Val-Leu-Lys-7-amido-4-methylcoumarin mutant enzyme W67L, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica
0.33
-
 Z-Val-Leu-Lys-7-amido-4-methylcoumarin wild type enzyme, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica
6
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin wild type enzyme, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica

General Information

General Information Comment Organism
physiological function the enzyme enables the parasite penetrate the intestinal wall Fasciola hepatica

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.59
-
 Z-Val-Leu-Lys-7-amido-4-methylcoumarin mutant enzyme W67L, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica
1.017
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme W67L, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica
1.037
-
 Z-Val-Leu-Lys-7-amido-4-methylcoumarin mutant enzyme H61N/W67L, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica
1.072
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme H61N/W67L, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica
20.76
-
 Z-Val-Leu-Lys-7-amido-4-methylcoumarin wild type enzyme, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica
1464
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin wild type enzyme, in 0.1 M sodium phosphate buffer, pH 6.0, 1 mM dithiothreitol and 1 mM EDTA at 25°C Fasciola hepatica