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Literature summary for 3.4.21.9 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
Search for more literature for 3.4.21.9

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
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

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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
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
0.3
-
 GDDDDK-2-naphthylamide mutant enzyme E136Y/R213L, at pH 8.3 and 30°C Oryzias latipes
0.4
-
 GDDDDK-2-naphthylamide mutant enzyme H24Q, at pH 8.3 and 30°C Oryzias latipes
0.5
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y/R213L, at pH 8.3 and 30°C Oryzias latipes
0.5
-
 GDDDDK-2-naphthylamide mutant enzyme R213L, at pH 8.3 and 30°C Oryzias latipes
0.6
-
 GDDDDK-2-naphthylamide mutant enzyme E136Y, at pH 8.3 and 30°C Oryzias latipes
0.6
-
 GDDDDK-2-naphthylamide wild type enzyme, at pH 8.3 and 30°C Oryzias latipes
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
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
0.9
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme R213L, at pH 8.3 and 30°C Oryzias latipes
1.1
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme H24Q, at pH 8.3 and 30°C Oryzias latipes
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
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
1.6
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y, at pH 8.3 and 30°C Oryzias latipes
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)

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

Organism

Organism UniProt Comment Textmining
Oryzias latipes A4UWM5 medaka
-

Purification (Commentary)

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

Renatured (Commentary)

Renatured (Comment) Organism
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
-
 Oryzias latipes ?
-
 ?
GDDDDK-2-naphthylamide + H2O specific substrate Oryzias latipes GDDDDK + 2-naphthylamine
-
 ?
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
-
 ?
trypsinogen + H2O recognition site DDDDK Oryzias latipes trypsin + ?
-
 ?

Synonyms

Synonyms Comment Organism
enterokinase
-
 Oryzias latipes
EPL enteropeptidase light chain Oryzias latipes

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
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
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
0.36
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y, at pH 8.3 and 30°C Oryzias latipes
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
0.56
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme R213L, at pH 8.3 and 30°C Oryzias latipes
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
0.64
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 8.3 and 30°C Oryzias latipes
1
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme H24Q, at pH 8.3 and 30°C Oryzias latipes
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
1.81
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y/R213L, at pH 8.3 and 30°C Oryzias latipes
17.8
-
 GDDDDK-2-naphthylamide mutant enzyme E136Y, at pH 8.3 and 30°C Oryzias latipes
26.2
-
 GDDDDK-2-naphthylamide mutant enzyme H24Q, at pH 8.3 and 30°C Oryzias latipes
27.9
-
 GDDDDK-2-naphthylamide wild type enzyme, at pH 8.3 and 30°C Oryzias latipes
28.75
-
 GDDDDK-2-naphthylamide mutant enzyme E136Y/R213L, at pH 8.3 and 30°C Oryzias latipes
33.02
-
 GDDDDK-2-naphthylamide mutant enzyme R213L, at pH 8.3 and 30°C Oryzias latipes

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.22
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y, at pH 8.3 and 30°C Oryzias latipes
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
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
0.41
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin wild type enzyme, at pH 8.3 and 30°C Oryzias latipes
0.62
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme R213L, at pH 8.3 and 30°C Oryzias latipes
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
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
0.89
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme H24Q, at pH 8.3 and 30°C Oryzias latipes
1.81
-
 benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin mutant enzyme E136Y/R213L, at pH 8.3 and 30°C Oryzias latipes
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
29.6
-
 GDDDDK-2-naphthylamide mutant enzyme E136Y, at pH 8.3 and 30°C Oryzias latipes
43.63
-
 GDDDDK-2-naphthylamide wild type enzyme, at pH 8.3 and 30°C Oryzias latipes
61.1
-
 GDDDDK-2-naphthylamide mutant enzyme R213L, at pH 8.3 and 30°C Oryzias latipes
63.1
-
 GDDDDK-2-naphthylamide mutant enzyme H24Q, at pH 8.3 and 30°C Oryzias latipes
99.5
-
 GDDDDK-2-naphthylamide mutant enzyme E136Y/R213L, at pH 8.3 and 30°C Oryzias latipes