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Literature summary extracted from

  • Jia, Y.; Quinn, C.M.; Bump, N.J.; Clark, K.M.; Clabbers, A.; Hardman, J.; Gagnon, A.; Kamens, J.; Tomlinson, M.J.; Wishart, N.; Allen, H.
    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.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.11.25 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

EC Number Protein Variants Comment Organism
2.7.11.25 D270A site-directed mutagenesis of COT30-397, inactive mutant Homo sapiens

General Stability

EC Number General Stability Organism
2.7.11.25 binding of p105 increases the solubility and stability of COT, but decreases catalytic activity Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.11.25 EGTA negative regulation of COT Homo sapiens
2.7.11.25 p105 inhibition of COT activity, but binding of p105 increases the solubility and stability of COT Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.11.25 0.00016
-
 MEK recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 5 mM Mn2+ Homo sapiens
2.7.11.25 0.00019
-
 MEK recombinant COT30-397, pH 7.5, 22°C, with 5 mM Mn2+ Homo sapiens
2.7.11.25 0.0003
-
 MEK recombinant COT30-397/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ Homo sapiens
2.7.11.25 0.0003
-
 MEK recombinant COT30-397/p105DELTAN, pH 7.5, 22°C, with 5 mM Mn2+ Homo sapiens
2.7.11.25 0.00031
-
 MEK recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ Homo sapiens
2.7.11.25 0.00036
-
 MEK recombinant COT30-397, pH 7.5, 22°C, with 10 mM Mg2+ Homo sapiens
2.7.11.25 0.02
-
 ATP recombinant COT30-397, pH 7.5, 22°C, with 5 mM Mn2+ Homo sapiens
2.7.11.25 0.03
-
 ATP recombinant COT30-397/p105DELTAN and recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 5 mM Mn2+ Homo sapiens
2.7.11.25 0.31
-
 ATP recombinant COT30-397/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ Homo sapiens
2.7.11.25 0.39
-
 ATP recombinant COT30-397, pH 7.5, 22°C, with 10 mM Mn2+ Homo sapiens
2.7.11.25 0.394
-
 ATP recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ Homo sapiens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.11.25 Mg2+
-
 Homo sapiens
2.7.11.25 Mn2+ preferred divalent cation as ATP metal cofactor Homo sapiens
2.7.11.25 additional information no or nearly no activity with Ni2+, Zn2+, Co2+, and Cu2+ Homo sapiens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.11.25 45000
-
 recombinant single COT30-397, gel filtration Homo sapiens
2.7.11.25 200000
-
 about, recombinant aggregated COT30-397, gel filtration Homo sapiens
2.7.11.25 600000
-
 about, recombinant aggregated, p105DELTAN-complexed COT30-467 Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.11.25 ATP + MEK Homo sapiens
-
 ADP + phosphorylated MEK
-
 ?
2.7.11.25 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

EC Number Organism UniProt Comment Textmining
2.7.11.25 Homo sapiens
-
-
-
2.7.12.2 Homo sapiens
-
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
2.7.12.2 phosphoprotein MEK is phosphorylated and activated by the mitogen-activated protein kinase kinase kinase COT, which can be inhibited by p105 Homo sapiens

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.11.25 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

EC Number Source Tissue Comment Organism Textmining
2.7.11.25 macrophage
-
 Homo sapiens
-
2.7.12.2 macrophage
-
 Homo sapiens
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.11.25 ATP + MEK
-
 Homo sapiens ADP + phosphorylated MEK
-
 ?
2.7.11.25 ATP + MEK substrate is a MAPKK, usage of inactivated GST-tagged mutant MEK Homo sapiens ADP + phosphorylated MEK
-
 ?
2.7.11.25 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

EC Number Subunits Comment Organism
2.7.11.25 More COT30-467 forms very tightly bound complexes with p105DELTAN which remain intact at 2-4 M urea Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
2.7.11.25 cancer osaka thyroid
-
 Homo sapiens
2.7.11.25 COT
-
 Homo sapiens
2.7.11.25 COT30-397
-
 Homo sapiens
2.7.11.25 COT30-467
-
 Homo sapiens
2.7.11.25 MAP3K
-
 Homo sapiens
2.7.11.25 MAPK kinase kinase
-
 Homo sapiens
2.7.12.2 ERK
-
 Homo sapiens
2.7.12.2 MEK
-
 Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.11.25 22
-
 assay at room temperature Homo sapiens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.11.25 0.005
-
 ATP recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ Homo sapiens
2.7.11.25 0.005
-
 MEK recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ Homo sapiens
2.7.11.25 0.006
-
 ATP recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 5 mM Mn2+ Homo sapiens
2.7.11.25 0.006
-
 MEK recombinant COT30-467/p105DELTAN, pH 7.5, 22°C, with 5 mM Mn2+ Homo sapiens
2.7.11.25 0.0328
-
 ATP recombinant COT30-397/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ Homo sapiens
2.7.11.25 0.0328
-
 MEK recombinant COT30-397/p105DELTAN, pH 7.5, 22°C, with 10 mM Mg2+ Homo sapiens
2.7.11.25 0.033
-
 ATP recombinant COT30-397/p105DELTAN, pH 7.5, 22°C, with 5 mM Mn2+ Homo sapiens
2.7.11.25 0.033
-
 MEK recombinant COT30-397/p105DELTAN, pH 7.5, 22°C, with 5 mM Mn2+ Homo sapiens
2.7.11.25 0.245
-
 MEK recombinant COT30-397, pH 7.5, 22°C, with 10 mM Mg2+ Homo sapiens
2.7.11.25 0.245
-
 ATP recombinant COT30-397, pH 7.5, 22°C, with 10 mM Mn2+ Homo sapiens
2.7.11.25 0.26
-
 ATP recombinant COT30-397, pH 7.5, 22°C, with 5 mM Mn2+ Homo sapiens
2.7.11.25 0.26
-
 MEK recombinant COT30-397, pH 7.5, 22°C, with 5 mM Mn2+ Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.11.25 7.5
-
 assay at Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.11.25 ATP
-
 Homo sapiens