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Literature summary for 2.7.11.25 extracted from
- Purification and kinetic characterization of recombinant human mitogen-activated protein kinase kinase kinase COT and the complexes with its cellular partner NF-kappa B1 p105 (2005), Arch. Biochem. Biophys., 441, 64-74.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression of FLAG-tagged COT30-467 and COT30-397 in a stable and soluble form when co-expressed with the C-terminal part of p105, complex formation with p105 reduces the kcat value of COT30-397 but increases the expression level, expression of wild-type and mutant COT30-397s in Spodoptera frugiperda Sf9 cells using the baculovirus infection system | Homo sapiens |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D270A | site-directed mutagenesis of COT30-397, inactive mutant | Homo sapiens |
General Stability
| General Stability | Organism |
|---|---|
| binding of p105 increases the solubility and stability of COT, but decreases catalytic activity | Homo sapiens |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| EGTA | negative regulation of COT | Homo sapiens |  |
| p105 | inhibition of COT activity, but binding of p105 increases the solubility and stability of COT | Homo sapiens |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.00016 | - |
MEK | recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 5 mM Mn2+ | Homo sapiens | |
| 0.00019 | - |
MEK | recombinant COT30-397, pH 7.5, 22°C, with 5 mM Mn2+ | Homo sapiens | |
| 0.0003 | - |
MEK | recombinant COT30-397/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ | Homo sapiens | |
| 0.0003 | - |
MEK | recombinant COT30-397/p105DELTAN, pH 7.5, 22°C, with 5 mM Mn2+ | Homo sapiens | |
| 0.00031 | - |
MEK | recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ | Homo sapiens | |
| 0.00036 | - |
MEK | recombinant COT30-397, pH 7.5, 22°C, with 10 mM Mg2+ | Homo sapiens | |
| 0.02 | - |
ATP | recombinant COT30-397, pH 7.5, 22°C, with 5 mM Mn2+ | Homo sapiens | |
| 0.03 | - |
ATP | recombinant COT30-397/p105DELTAN and recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 5 mM Mn2+ | Homo sapiens | |
| 0.31 | - |
ATP | recombinant COT30-397/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ | Homo sapiens | |
| 0.39 | - |
ATP | recombinant COT30-397, pH 7.5, 22°C, with 10 mM Mn2+ | Homo sapiens | |
| 0.394 | - |
ATP | recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ | Homo sapiens | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | - |
Homo sapiens | |
| Mn2+ | preferred divalent cation as ATP metal cofactor | Homo sapiens | |
| additional information | no or nearly no activity with Ni2+, Zn2+, Co2+, and Cu2+ | Homo sapiens |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 45000 | - |
recombinant single COT30-397, gel filtration | Homo sapiens |
| 200000 | - |
about, recombinant aggregated COT30-397, gel filtration | Homo sapiens |
| 600000 | - |
about, recombinant aggregated, p105DELTAN-complexed COT30-467 | Homo sapiens |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + MEK | Homo sapiens | - |
ADP + phosphorylated MEK | - |
? | |
| additional information | Homo sapiens | COT is a proto-oncogene, the enzyme is essential for the lipopolysaccharide activation of the ERK MAPK cascade in macrophages | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Homo sapiens | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| purification of recombinant wild-type and mutant COT30-397s, free or complexed to p105DELTAN, and of p105DELTAN-complexed recombinant COT30-467 from Sf9 insect cells by anti-Flag immunoaffinity chromatography, the purification of free COT30-467 is not possible due to its extremely low expression level in absence of p105DELTAN | Homo sapiens |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| macrophage | - |
Homo sapiens | - |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + MEK | - |
Homo sapiens | ADP + phosphorylated MEK | - |
? | |
| ATP + MEK | substrate is a MAPKK, usage of inactivated GST-tagged mutant MEK | Homo sapiens | ADP + phosphorylated MEK | - |
? | |
| additional information | COT is a proto-oncogene, the enzyme is essential for the lipopolysaccharide activation of the ERK MAPK cascade in macrophages | Homo sapiens | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| More | COT30-467 forms very tightly bound complexes with p105DELTAN which remain intact at 2-4 M urea | Homo sapiens |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| cancer osaka thyroid | - |
Homo sapiens |
| COT | - |
Homo sapiens |
| COT30-397 | - |
Homo sapiens |
| COT30-467 | - |
Homo sapiens |
| MAP3K | - |
Homo sapiens |
| MAPK kinase kinase | - |
Homo sapiens |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 22 | - |
assay at room temperature | Homo sapiens |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.005 | - |
ATP | recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ | Homo sapiens | |
| 0.005 | - |
MEK | recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ | Homo sapiens | |
| 0.006 | - |
ATP | recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 5 mM Mn2+ | Homo sapiens | |
| 0.006 | - |
MEK | recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 5 mM Mn2+ | Homo sapiens | |
| 0.0328 | - |
ATP | recombinant COT30-397/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ | Homo sapiens | |
| 0.0328 | - |
MEK | recombinant COT30-397/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ | Homo sapiens | |
| 0.033 | - |
ATP | recombinant COT30-397/p105DELTAN, pH 7.5, 22°C, with 5 mM Mn2+ | Homo sapiens | |
| 0.033 | - |
MEK | recombinant COT30-397/p105DELTAN, pH 7.5, 22°C, with 5 mM Mn2+ | Homo sapiens | |
| 0.245 | - |
MEK | recombinant COT30-397, pH 7.5, 22°C, with 10 mM Mg2+ | Homo sapiens | |
| 0.245 | - |
ATP | recombinant COT30-397, pH 7.5, 22°C, with 10 mM Mn2+ | Homo sapiens | |
| 0.26 | - |
ATP | recombinant COT30-397, pH 7.5, 22°C, with 5 mM Mn2+ | Homo sapiens | |
| 0.26 | - |
MEK | recombinant COT30-397, pH 7.5, 22°C, with 5 mM Mn2+ | Homo sapiens |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
assay at | Homo sapiens |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| ATP | - |
Homo sapiens | |
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