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Literature summary for 3.4.22.59 extracted from

  • Velazquez-Delgado, E.M.; Hardy, J.A.
    Zinc-mediated allosteric inhibition of caspase-6 (2012), J. Biol. Chem., 287, 36000-36011.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.22.59

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Homo sapiens

Crystallization (Commentary)

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

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

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 [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

Organism UniProt Comment Textmining
Homo sapiens P55212
-
-

Purification (Commentary)

Purification (Comment) Organism
Hi-Trap nickel-affinity column chromatography Homo sapiens

Substrates and Products (Substrate)

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

Synonyms Comment Organism
Casp-6
-
 Homo sapiens

Turnover Number [1/s]

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 [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 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 [mM/s]

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