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Literature summary extracted from

  • Xu, J.; Hu, S.; Wang, X.; Zhao, Z.; Zhang, X.; Wang, H.; Zhang, D.; Guo, Y.
    Structure basis for the unique specificity of medaka enteropeptidase light chain (2014), Protein Cell, 5, 178-181.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.21.9 expressed in Escherichia coli BL21(DE3) cells Oryzias latipes

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.4.21.9 hanging drop vapor diffusion method, using 20% (w/v) polyethylene glycol 3350, 0.1 M cadmium chloride, 0.1 M sodium acetate (pH 4.6) Oryzias latipes

Protein Variants

EC Number Protein Variants Comment Organism
3.4.21.9 E136Y the mutant shows clearly reduced activity compared to the wild type enzyme Oryzias latipes
3.4.21.9 E136Y/R213L the mutant shows about wild type activity Oryzias latipes
3.4.21.9 H24Q the mutant shows slightly reduced activity compared to the wild type enzyme Oryzias latipes
3.4.21.9 R213L the mutant shows increased activity compared to the wild type enzyme Oryzias latipes

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.21.9 0.2
-
 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y/R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.3
-
 GDDDDK-2-naphthylamide mutant enzyme E136Y/R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.4
-
 GDDDDK-2-naphthylamide mutant enzyme H24Q, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.5
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y/R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.5
-
 GDDDDK-2-naphthylamide mutant enzyme R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.6
-
 GDDDDK-2-naphthylamide mutant enzyme E136Y, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.6
-
 GDDDDK-2-naphthylamide wild type enzyme, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.7
-
 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin mutant enzyme H24Q, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.8
-
 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin mutant enzyme R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.9
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 1.1
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme H24Q, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 1.3
-
 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 1.5
-
 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 1.6
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 1.6
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 8.3 and 30°C Oryzias latipes

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.4.21.9 trypsinogen + H2O Oryzias latipes recognition site DDDDK trypsin + ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.4.21.9 Oryzias latipes A4UWM5 medaka
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.21.9 STI-agarose column chromatography Oryzias latipes

Renatured (Commentary)

EC Number Renatured (Comment) Organism
3.4.21.9 fast dilution method in 20 mM NH3 and 10% (v/v) glycerol at pH 10.5 for 72 h at 4? Oryzias latipes

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.21.9 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
-
 Oryzias latipes ?
-
 ?
3.4.21.9 GDDDDK-2-naphthylamide + H2O specific substrate Oryzias latipes GDDDDK + 2-naphthylamine
-
 ?
3.4.21.9 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin + H2O
-
 Oryzias latipes t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg + 7-amino-4-methylcoumarin
-
 ?
3.4.21.9 trypsinogen + H2O recognition site DDDDK Oryzias latipes trypsin + ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.4.21.9 enterokinase
-
 Oryzias latipes
3.4.21.9 EPL enteropeptidase light chain Oryzias latipes

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.21.9 0.3
-
 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.34
-
 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.36
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.53
-
 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin mutant enzyme R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.56
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.6
-
 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin mutant enzyme H24Q, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.64
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 1
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme H24Q, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 1.38
-
 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y/R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 1.81
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y/R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 17.8
-
 GDDDDK-2-naphthylamide mutant enzyme E136Y, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 26.2
-
 GDDDDK-2-naphthylamide mutant enzyme H24Q, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 27.9
-
 GDDDDK-2-naphthylamide wild type enzyme, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 28.75
-
 GDDDDK-2-naphthylamide mutant enzyme E136Y/R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 33.02
-
 GDDDDK-2-naphthylamide mutant enzyme R213L, at pH 8.3 and 30°C Oryzias latipes

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.4.21.9 0.22
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.23
-
 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.23
-
 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.41
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.62
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.67
-
 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin mutant enzyme R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.85
-
 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin mutant enzyme H24Q, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 0.89
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme H24Q, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 1.81
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y/R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 2.29
-
 t-butyloxycarbonyl-E(benzyl ester)-Ala-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y/R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 29.6
-
 GDDDDK-2-naphthylamide mutant enzyme E136Y, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 43.63
-
 GDDDDK-2-naphthylamide wild type enzyme, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 61.1
-
 GDDDDK-2-naphthylamide mutant enzyme R213L, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 63.1
-
 GDDDDK-2-naphthylamide mutant enzyme H24Q, at pH 8.3 and 30°C Oryzias latipes
3.4.21.9 99.5
-
 GDDDDK-2-naphthylamide mutant enzyme E136Y/R213L, at pH 8.3 and 30°C Oryzias latipes