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Literature summary for 3.4.21.9 extracted from

  • Smith, E.T.; Johnson, D.A.
    Human enteropeptidase light chain: bioengineering of recombinants and kinetic investigations of structure and function (2013), Protein Sci., 22, 577-585.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Pichia pastoris X-33 strain Homo sapiens

Protein Variants

Protein Variants Comment Organism
R96Q the mutant shows decreased specificities for substrates containing the sequences DDDDK and DDDDR, while basic pancreatic trypsin inhibitor inhibition is increased Homo sapiens
Y174R the mutant shows improved specificities for substrates containing the sequences DDDDK and DDDDR, while basic pancreatic trypsin inhibitor inhibition is significantly decreased Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
basic pancreatic trypsin inhibitor reversible inhibition Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00258
-
GD4R-4-nitroanilide mutant enzyme Y174R, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
0.00629
-
GD4R-4-nitroanilide wild type enzyme, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
0.0179
-
GDDDDK-4-nitroanilide mutant enzyme Y174R, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
0.018
-
GD4R-4-nitroanilide mutant enzyme R96Q, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
0.0298
-
GDDDDK-4-nitroanilide wild type enzyme, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
0.165
-
Z-Lys-SBzl mutant enzyme R96Q, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
0.172
-
Z-Lys-SBzl mutant enzyme Y174R, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
0.19
-
Z-Lys-SBzl wild type enzyme, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
0.276
-
GDDDDK-4-nitroanilide mutant enzyme R96Q, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ CaCl2 enhances enzyme activity at 0.02 mM Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
26000
-
x * 26000, recombinant catalytic subunit, SDS-PAGE Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
trypsinogen + H2O Homo sapiens cleavage sequence DDDDK-X trypsin + ?
-
?

Organism

Organism UniProt Comment Textmining
Homo sapiens P98073
-
-

Purification (Commentary)

Purification (Comment) Organism
soybean trypsin inhibitor-agarose column chromatography, and gel filtration Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
GD4R-4-nitroanilide + H2O
-
Homo sapiens ?
-
?
GDDDDK-4-nitroanilide + H2O
-
Homo sapiens GDDDDK + 4-nitroaniline
-
?
trypsinogen + H2O cleavage sequence DDDDK-X Homo sapiens trypsin + ?
-
?
Z-Lys-SBzl + H2O
-
Homo sapiens ?
-
?

Subunits

Subunits Comment Organism
? x * 26000, recombinant catalytic subunit, SDS-PAGE Homo sapiens

Synonyms

Synonyms Comment Organism
enterokinase
-
Homo sapiens
EPL catalytic subunit Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
42
-
GD4R-4-nitroanilide mutant enzyme R96Q, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
45
-
GD4R-4-nitroanilide wild type enzyme, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
49
-
GD4R-4-nitroanilide mutant enzyme Y174R, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
122
-
GDDDDK-4-nitroanilide mutant enzyme Y174R, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
135
-
GDDDDK-4-nitroanilide wild type enzyme, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
148
-
GDDDDK-4-nitroanilide mutant enzyme R96Q, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
319
-
Z-Lys-SBzl mutant enzyme Y174R, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
336
-
Z-Lys-SBzl wild type enzyme, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
354
-
Z-Lys-SBzl mutant enzyme R96Q, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0000461
-
basic pancreatic trypsin inhibitor wild type enzyme, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
0.0000632
-
basic pancreatic trypsin inhibitor mutant enzyme R96Q, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
0.000228
-
basic pancreatic trypsin inhibitor mutant enzyme Y174R, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
537
-
GDDDDK-4-nitroanilide mutant enzyme R96Q, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
1770
-
Z-Lys-SBzl wild type enzyme, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
1860
-
Z-Lys-SBzl mutant enzyme Y174R, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
2150
-
Z-Lys-SBzl mutant enzyme R96Q, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
2330
-
GD4R-4-nitroanilide mutant enzyme R96Q, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
4530
-
GDDDDK-4-nitroanilide wild type enzyme, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
6830
-
GDDDDK-4-nitroanilide mutant enzyme Y174R, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
7200
-
GD4R-4-nitroanilide wild type enzyme, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens
18900
-
GD4R-4-nitroanilide mutant enzyme Y174R, in 100 mM Tris, 0.02 mM CaCl2, 0.01% (v/v) Triton X-100, 10% (v/v) DMSO, pH 8.0, at 22°C Homo sapiens