Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain Bl21 (DE3) pLysS | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
wild-type and mutant caspase-2 enzymes complexed with peptide aldehyde inhibitors, incubation of 3 mg/ml protein with 1-5 mM peptide aldehyde inhibitor in DMSO at room temperature for 2 h, hanging drop vapour diffusion method, mixing of 0.002 ml of protein complex solution with 0.002 ml of reservoir solution containing 0.1 M HEPES, pH 7.0, 15% PEG 3350, 3 mM DTT, X-ray diffraction structure determination and analysis at 1.5-2.4 A resolution, molecular replacement | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
T380A | site-directed mutagenesis | Homo sapiens |
T380A/Y420A | site-directed mutagenesis | Homo sapiens |
Y420A | site-directed mutagenesis | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ac-ADVAD-CHO | binding structure | Homo sapiens | |
Ac-DVAD-CHO | binding structure | Homo sapiens | |
Ac-VDVAD-CHO | binding structure | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.025 | - |
Ac-VDVAD-7-amido-4-trifluoromethylcoumarin | pH 6.5, 22°C, wild-type enzyme | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P42575 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
proteolytic modification | caspase-2 undergoes autocatalytic activation to remove the prodomain and linker region to generate a stable dimer consisting of the large subunit p19 and the small subunit p12. This p19/p12 dimer self-associates to form the active caspase-2, forming a dimer, a tetramer, or a dimer-of-dimers | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Ac-VDVAD-7-amido-4-trifluoromethylcoumarin + H2O | - |
Homo sapiens | Ac-VDVAD + 7-amido-4-trifluoromethylcoumarin | - |
? | |
additional information | caspase-2 protein substrate recognition and catalysis, structural basis for the essential P5 recognition of caspase-2, molecular replacement and modeling using the structure with PDB ID 1PYO, overview. The key residues involved in catalysis, including the catalytic dyad, formed by Thr380 and Tyr420, and the invariant Arg378, superpose with each other regardless of the peptide inhibitor binding. Caspase-2 uniquely prefers a pentapeptide, such as VDVAD, rather than a tetrapeptide, as required for efficient cleavage by other caspases. The P3 Val is only involved in main chain hydrogen bonds with Arg378, and its side chain makes no contacts with the enzyme. The nitrogen atom of P4 Asp forms a hydrogen bond with the carbonyl oxygen of Tyr420, and its side chain extensively interacts with the enzyme through hydrogen bonds with the backbone of Tyr420 and the side chains of Trp385, Asn379, Arg417, and Glu418 | Homo sapiens | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
oligomer | caspase-2 undergoes autocatalytic activation to remove the prodomain and linker region to generate a stable dimer consisting of the large subunit p19, residues 170-333, and the small subunit p12, residues 348-452. This p19/p12 dimer self-associates to form the active caspase-2 | Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at room temperature | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.6 | - |
Ac-VDVAD-7-amido-4-trifluoromethylcoumarin | pH 6.5, 22°C, wild-type enzyme | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
assay at | Homo sapiens |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.000025 | - |
pH 6.5, 22°C, wild-type enzyme | Homo sapiens | Ac-VDVAD-CHO | |
0.00011 | - |
pH 6.5, 22°C, wild-type enzyme | Homo sapiens | Ac-ADVAD-CHO | |
0.00071 | - |
pH 6.5, 22°C, wild-type enzyme | Homo sapiens | Ac-DVAD-CHO |
General Information | Comment | Organism |
---|---|---|
evolution | caspase-2 is the most evolutionarily conserved member in the human caspase family | Homo sapiens |
additional information | caspase-2 is synthesized as an inactive zymogen. The zymogen sequence includes a long prodomain containing a CARD followed by a large domain, a linker, and a small domain. Caspase-2 undergoes autocatalytic activation to remove the prodomain and linker region to generate a stable dimer consisting of the large subunit p19 and the small subunit p12. This p19/p12 dimer self-associates to form the active caspase-2 | Homo sapiens |
physiological function | caspase-2 plays important roles in stress-induced apoptosis, cell cycle regulation, and tumor suppression | Homo sapiens |