Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Huber, K.L.; Hardy, J.A.
    Mechanism of zinc-mediated inhibition of caspase-9 (2012), Protein Sci., 21, 1056-1065.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.4.22.62 additional information the uncleaved monomeric zymogen of caspase-9 has very low activity, which is increased upon dimerization. In dimeric caspase-9 cleavage at a specific aspartate residue in the intersubuint linker between the large and small subunits is required for caspase-9 to attain increased activity. Full enzymatic activity is achieved only upon interaction with the apoptosome, a multicomponent, heptameric caspase-9 activating platform Homo sapiens

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.22.62 expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain B21(DE3) in incusion bodies Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
3.4.22.62 C172A the mutant enzyme shows reduced zinc binding compared to the wild-type enzyme Homo sapiens
3.4.22.62 C239S the mutant enzyme shows reduced zinc binding compared to the wild-type enzyme Homo sapiens
3.4.22.62 C272A the mutant enzyme shows reduced zinc binding compared to the wild-type enzyme Homo sapiens
3.4.22.62 C272A/C287A the mutant enzyme shows highly reduced zinc binding compared to the wild-type enzyme Homo sapiens
3.4.22.62 C287A the active site mutant enzyme shows reduced zinc binding compared to the wild-type enzyme Homo sapiens
3.4.22.62 C287A/C239S the mutant enzyme shows reduced zinc binding compared to the wild-type enzyme Homo sapiens
3.4.22.62 H237A the active site mutant enzyme shows reduced zinc binding compared to the wild-type enzyme Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.22.62 Mn2+ slight inhibition Homo sapiens
3.4.22.62 additional information no inhibition by Co2+, PB2+, and Cd2+ Homo sapiens
3.4.22.62 NO3- slight inhibition Homo sapiens
3.4.22.62 Zn2+ mixed-tpe inhibition, kinetics and mechanism of zinc-mediated inhibition of caspase-9, overview. Two distinct zinc-binding sites on caspase-9, the first site, composed of H237, C239, and C287, includes the active site dyad and is primarily responsible for zinc-mediated inhibition. The second binding site at C272 is distal from the active site. EDTA can hinder enzyme inhibition by Zn2+. Zinc-mediated inhibition does not influence the overall structure of caspase-9 monomers. Each wild-type caspase-9 monomer binds two zinc ions. Caspase-9 variants in which the active-site residues are replaced, C287A or H237A, bind just one zinc Homo sapiens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.4.22.62 Cd2+ slight activation Homo sapiens
3.4.22.62 Co2+ slight activation Homo sapiens
3.4.22.62 Pb2+ slight activation Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
3.4.22.62 Homo sapiens
-
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.4.22.62 proteolytic modification the uncleaved monomeric zymogen of caspase-9 has very low activity, which is increased upon dimerization. In dimeric caspase-9 cleavage at a specific aspartate residue in the intersubuint linker between the large and small subunits is required for caspase-9 to attain increased activity Homo sapiens

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.22.62 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain B21(DE3) inclusion bodies by solubilization with guanidine hydrochloride and dialysis, followed by nickel affinity and anion exchange chromatography Homo sapiens

Renatured (Commentary)

EC Number Renatured (Comment) Organism
3.4.22.62 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain B21(DE3) incusion bodies by solubilization with guanidine hydrochloride and dialysis Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.22.62 LEHD-7-amido-4-methylcoumarin + H2O
-
 Homo sapiens LEHD + 7-amino-4-methylcoumarin
-
 ?

Subunits

EC Number Subunits Comment Organism
3.4.22.62 homodimer gel filtration and SDS-PAGE, caspases are functional as homodimers of monomeric units that comprise an N-terminal prodomain and a catalytic large and small subunit connected by an intersubunit linker. The zymogen (uncleaved) caspase-9 as a monomer has very low activity, which is increased upon dimerization Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.4.22.62 37
-
 assay at Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4.22.62 6.5
-
 assay at Homo sapiens

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.4.22.62 0.0002
-
 Zn2+ mutant C272A/C287A, pH 6.5, 37°C Homo sapiens
3.4.22.62 0.0007
-
 Zn2+ mutant C272A, pH 6.5, 37°C Homo sapiens
3.4.22.62 0.0007
-
 Zn2+ mutant C287A/C239S, pH 6.5, 37°C Homo sapiens
3.4.22.62 0.0008
-
 Zn2+ mutant C287A, pH 6.5, 37°C Homo sapiens
3.4.22.62 0.0008
-
 Zn2+ mutant H237A, pH 6.5, 37°C Homo sapiens
3.4.22.62 0.0012
-
 Zn2+ mutant C239S, pH 6.5, 37°C Homo sapiens
3.4.22.62 0.0015
-
 Zn2+ mutant C172A, pH 6.5, 37°C Homo sapiens
3.4.22.62 0.0018
-
 Zn2+ wild-type enzyme, pH 6.5, 37°C Homo sapiens

General Information

EC Number General Information Comment Organism
3.4.22.62 physiological function initiator caspases, such as caspase-9, function in recruitment and regulation and are designated as caspase activation and recruitment domains. As an initiator, caspase-9 regulates the upstream stages of the apoptotic caspase cascade, making it a critical control point. Inhibition by zinc has a regulatory function, and its relief is central to both small-molecule and natively induced caspase activation Homo sapiens