Literature summary for 3.4.22.B61 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.

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Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Saccharomyces cerevisiae	Fasciola hepatica

Protein Variants

Protein Variants	Comment	Organism
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	Fasciola hepatica

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.02481	-	benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin	wild type enzyme, at pH 5.5 and 37°C	Fasciola hepatica
0.05392	-	tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin	wild type enzyme, at pH 5.5 and 37°C	Fasciola hepatica
0.0786	-	tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin	mutant enzyme L69W, at pH 5.5 and 37°C	Fasciola hepatica
0.1373	-	benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin	mutant enzyme L69W, at pH 5.5 and 37°C	Fasciola hepatica

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
38000	-	x * 38000, proenzyme, SDS-PAGE	Fasciola hepatica

Organism

Organism	UniProt	Comment	Textmining
Fasciola hepatica	Q9NB30	-	-

Purification (Commentary)

Purification (Comment)	Organism
nickel-chelating Sepharose column chromatography and Mono S column chromatography	Fasciola hepatica

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
Albumin + H2O	weak substrate	Fasciola hepatica	?	-	?
benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin + H2O	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	Fasciola hepatica	benzyloxycarbonyl-L-Phe-L-Arg + 7-amino-4-methylcoumarin	-	?
casein + H2O	-	Fasciola hepatica	?	-	?
Hemoglobin + H2O	cathepsin l5 hydrolyzes hemoglobin only at pH 5.5	Fasciola hepatica	?	-	?
Immunoglobulin + H2O	-	Fasciola hepatica	?	-	?
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	Fasciola hepatica	?	-	?
tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin + H2O	weak substrate	Fasciola hepatica	tosyl-Gly-L-Pro-L-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	Fasciola hepatica	?	-	?
type 4 collagen + H2O	-	Fasciola hepatica	?	-	?

Subunits

Subunits	Comment	Organism
?	x * 38000, proenzyme, SDS-PAGE	Fasciola hepatica

Synonyms

Synonyms	Comment	Organism
CatL5	-	Fasciola hepatica

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.84	-	tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin	wild type enzyme, at pH 5.5 and 37°C	Fasciola hepatica
1.34	-	tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin	mutant enzyme L69W, at pH 5.5 and 37°C	Fasciola hepatica
11.05	-	benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin	mutant enzyme L69W, at pH 5.5 and 37°C	Fasciola hepatica
17.89	-	benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin	wild type enzyme, at pH 5.5 and 37°C	Fasciola hepatica

pH Optimum

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

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
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	Fasciola hepatica

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
15.5	-	tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin	wild type enzyme, at pH 5.5 and 37°C	Fasciola hepatica
17	-	tosyl-Gly-L-Pro-L-Arg-7-amido-4-methylcoumarin	mutant enzyme L69W, at pH 5.5 and 37°C	Fasciola hepatica
80.5	-	benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin	mutant enzyme L69W, at pH 5.5 and 37°C	Fasciola hepatica
721	-	benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin	wild type enzyme, at pH 5.5 and 37°C	Fasciola hepatica

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Release 2023.1 (January 2023)