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Literature summary for 3.4.22.B61 extracted from

  • 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 (2012), Biochimie, 94, 1119-1127.
    View publication on PubMed
Search for more literature for 3.4.22.B61

Cloned(Commentary)

Cloned (Comment) Organism
expressed in yeast Fasciola hepatica

Protein Variants

Protein Variants Comment Organism
L69C the mutation results in low overall activity Fasciola hepatica
L69F the mutant shows a 2-3fold increase in the ability to cleave substrates with P2 proline Fasciola hepatica
L69S the mutation results in low overall activity Fasciola hepatica
L69Y the mutant shows a 2-3fold increase in the ability to cleave substrates with P2 proline Fasciola hepatica

Inhibitors

Inhibitors Comment Organism Structure
E-64
-
 Fasciola hepatica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.01272
-
 Z-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme L69S, at pH 5.5 and 37°C Fasciola hepatica
0.01372
-
 Z-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme L69C, at pH 5.5 and 37°C Fasciola hepatica
0.01911
-
 Z-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme L69F, at pH 5.5 and 37°C Fasciola hepatica
0.02481
-
 Z-Phe-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5 and 37°C Fasciola hepatica
0.03287
-
 Z-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme L69Y, at pH 5.5 and 37°C Fasciola hepatica
0.05193
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme L69Y, at pH 5.5 and 37°C Fasciola hepatica
0.06346
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme L69Y, at pH 5.5 and 37°C Fasciola hepatica
0.07674
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme L69C, at pH 5.5 and 37°C Fasciola hepatica
0.1269
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme L69S, at pH 5.5 and 37°C Fasciola hepatica
5392
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5 and 37°C Fasciola hepatica

Organism

Organism UniProt Comment Textmining
Fasciola hepatica Q9NB30
-
-

Purification (Commentary)

Purification (Comment) Organism
nickel chelating column chromatography and Mono S column chromatography 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
-
 ?
Z-Phe-Arg-7-amido-4-methylcoumarin + H2O preferred substrate Fasciola hepatica Z-Phe-Arg + 7-amino-4-methylcoumarin
-
 ?

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
0.19
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme L69C, at pH 5.5 and 37°C Fasciola hepatica
0.27
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme L69S, at pH 5.5 and 37°C Fasciola hepatica
0.84
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5 and 37°C Fasciola hepatica
1.73
-
 Z-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme L69S, at pH 5.5 and 37°C Fasciola hepatica
1.78
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme L69Y, at pH 5.5 and 37°C Fasciola hepatica
1.92
-
 Z-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme L69C, at pH 5.5 and 37°C Fasciola hepatica
2.54
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme L69Y, at pH 5.5 and 37°C Fasciola hepatica
17.89
-
 Z-Phe-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5 and 37°C Fasciola hepatica
17.93
-
 Z-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme L69F, at pH 5.5 and 37°C Fasciola hepatica
21.52
-
 Z-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme L69Y, at pH 5.5 and 37°C Fasciola hepatica

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.1
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme L69S, at pH 5.5 and 37°C Fasciola hepatica
2.5
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme L69C, at pH 5.5 and 37°C Fasciola hepatica
15.5
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5 and 37°C Fasciola hepatica
34.4
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme L69Y, at pH 5.5 and 37°C Fasciola hepatica
40
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin mutant enzyme L69Y, at pH 5.5 and 37°C Fasciola hepatica
136
-
 Z-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme L69S, at pH 5.5 and 37°C Fasciola hepatica
140
-
 Z-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme L69C, at pH 5.5 and 37°C Fasciola hepatica
655
-
 Z-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme L69Y, at pH 5.5 and 37°C Fasciola hepatica
721
-
 Z-Phe-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 5.5 and 37°C Fasciola hepatica
938
-
 Z-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme L69F, at pH 5.5 and 37°C Fasciola hepatica