Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapor diffusion method, using 0.1 M sodium acetate, pH 4.6, 2 M NaCl, and 3% (v/v) ethanol | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
C163S | the mutant is catalytically inactive and not capable of self-cleavage | Homo sapiens |
E244A | the function of the mutant is crippled by 2fold compared to the wild type enzyme | Homo sapiens |
E244A/H287A | the function of the mutant is crippled by 47fold compared to the wild type enzyme | Homo sapiens |
H287A | the function of the mutant is crippled by 14fold compared to the wild type enzyme | Homo sapiens |
K36A | the function of the mutant is crippled by 2fold compared to the wild type enzyme | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | caspase-6 is allosterically inhibited by zinc and binds one zinc/monomer at an exosite distal from the active site. Complete inhibition at 0.0015 mM | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.082 | - |
N-acetyl-VEID-7-amido-4-methylcoumarin | mutant enzyme E244A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
0.093 | - |
N-acetyl-VEID-7-amido-4-methylcoumarin | wild type enzyme, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
0.109 | - |
N-acetyl-VEID-7-amido-4-methylcoumarin | mutant enzyme K36A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
0.14 | - |
N-acetyl-VEID-7-amido-4-methylcoumarin | mutant enzyme H287A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
0.176 | - |
N-acetyl-VEID-7-amido-4-methylcoumarin | mutant enzyme E244A/H287A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P55212 | - |
- |
Purification (Comment) | Organism |
---|---|
Hi-Trap nickel-affinity column chromatography | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
N-acetyl-VEID-7-amido-4-methylcoumarin + H2O | - |
Homo sapiens | N-acetyl-VEID + 7-amino-4-methylcoumarin | - |
? | |
procaspase-6 mutant C163S + H2O | cleavage at the intersubunit linker position Asp-193 | Homo sapiens | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Casp-6 | - |
Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.015 | - |
N-acetyl-VEID-7-amido-4-methylcoumarin | mutant enzyme E244A/H287A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
0.051 | - |
N-acetyl-VEID-7-amido-4-methylcoumarin | mutant enzyme E244A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
0.309 | - |
N-acetyl-VEID-7-amido-4-methylcoumarin | mutant enzyme H287A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
0.371 | - |
N-acetyl-VEID-7-amido-4-methylcoumarin | mutant enzyme K36A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
0.7 | - |
N-acetyl-VEID-7-amido-4-methylcoumarin | wild type enzyme, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00015 | - |
Zn2+ | wild type enzyme, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
0.003 | - |
Zn2+ | mutant enzyme E244A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
0.0046 | - |
Zn2+ | mutant enzyme K36A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
0.007 | - |
Zn2+ | mutant enzyme H287A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
0.0072 | - |
Zn2+ | mutant enzyme E244A/H287A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.0034 | - |
wild type enzyme, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | Zn2+ | |
0.0107 | - |
mutant enzyme E244A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | Zn2+ | |
0.0158 | - |
mutant enzyme K36A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | Zn2+ | |
0.0249 | - |
mutant enzyme H287A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | Zn2+ | |
0.0256 | - |
mutant enzyme E244A/H287A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | Zn2+ |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
100 | - |
N-acetyl-VEID-7-amido-4-methylcoumarin | mutant enzyme E244A/H287A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
600 | - |
N-acetyl-VEID-7-amido-4-methylcoumarin | mutant enzyme E244A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
2200 | - |
N-acetyl-VEID-7-amido-4-methylcoumarin | mutant enzyme H287A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
3400 | - |
N-acetyl-VEID-7-amido-4-methylcoumarin | mutant enzyme K36A, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens | |
7500 | - |
N-acetyl-VEID-7-amido-4-methylcoumarin | wild type enzyme, in 100 mM HEPES, pH 7.5, 10% (w/v) sucrose, 0.1% (w/v) CHAPS, 20 mM TCEP, and 120 mM NaCl, at 37°C | Homo sapiens |