Application | Comment | Organism |
---|---|---|
analysis | development of a continuous spectrophotometric assay for mitogen-activated protein kinase kinases | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
S207E/T211E | a constitutively active MKK6 mutant | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Homo sapiens | |
0.000129 | - |
K52R-[ERK2] | pH 7.0, 25°C | Homo sapiens | |
0.00016 | - |
K53M-[p38alpha] | pH 7.0, 25°C | Homo sapiens | |
0.000184 | - |
ERK2 | pH 7.0, 25°C | Homo sapiens | |
0.000205 | - |
p38alpha | pH 7.0, 25°C | Homo sapiens | |
0.0533 | - |
ATP | pH 7.0, 25°C | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + a protein | Homo sapiens | - |
ADP + a phosphoprotein | - |
? | |
ATP + ERK2 | Homo sapiens | - |
ADP + phosphorylated ERK2 | - |
? | |
ATP + p38alpha | Homo sapiens | MKK6 phosphorylates p38 MAPK on Thr180 and Tyr182, the sites of phosphorylation that activate p38 MAPK | ADP + phosphorylated p38alpha | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P52564 | - |
- |
Homo sapiens | Q02750 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + a protein | - |
Homo sapiens | ADP + a phosphoprotein | - |
? | |
ATP + ERK2 | - |
Homo sapiens | ADP + phosphorylated ERK2 | - |
? | |
ATP + ERK2 | MEK1 is able to effectively phosphorylate Thr183 and Tyr185 in the activation loop of ERK2 | Homo sapiens | ADP + phosphorylated ERK2 | - |
? | |
ATP + K52R-[ERK2] | catalytically inactive ERK2 in which lysine-52 is substituted with arginine | Homo sapiens | ADP + phospho-K52R-[ERK2] | - |
? | |
ATP + K53M-[p38alpha] | catalytically inactive p38alpha in which lysine-53 is substituted with methionine | Homo sapiens | ADP + phospho-K53M-[p38alpha] | - |
? | |
ATP + p38alpha | MKK6 phosphorylates p38 MAPK on Thr180 and Tyr182, the sites of phosphorylation that activate p38 MAPK | Homo sapiens | ADP + phosphorylated p38alpha | - |
? | |
additional information | development of a continuous spectrophotometric assay for mitogen-activated protein kinase kinases. The assay relies on the measurement of phosphoprotein product generated in the first step of the MAPK kinase reaction. Dephosphorylation of the phosphoprotein is coupled to a MAPK phosphatase, MKP3, to generate phosphate, which is then used as the substrate of purine nucleoside phosphorylase to catalyze the N-glycosidic cleavage of 2-amino 6-mercapto 7-methyl purine ribonucleoside. Of the reaction products ribose 1-phosphate and 2-amino 6-mercapto 7-methylpurine, the latter has a high absorbance at 360 nm relative to the nucleoside and, hence, provides a spectrophotometric signal that can be continuously followed. In the presence of excess phosphatase, the phosphorylated protein substrate molecules undergo dephosphorylation almost immediately after their formation. The steady-state use of the resultant inorganic phosphate is a reflection of the constant initial velocity of the exchange reaction. Method development and evaluation, overview | Homo sapiens | ? | - |
? | |
additional information | development of a continuous spectrophotometric assay for mitogen-activated protein kinase kinases. The assay relies on the measurement of phosphoprotein product generated in the first step of the MAPK kinase reaction. Dephosphorylation of the phosphoprotein is coupled to a MAPK phosphatase, MKP5, to generate phosphate, which is then used as the substrate of purine nucleoside phosphorylase to catalyze the N-glycosidic cleavage of 2-amino 6-mercapto 7-methyl purine ribonucleoside. Of the reaction products ribose 1-phosphate and 2-amino 6-mercapto 7-methylpurine, the latter has a high absorbance at 360 nm relative to the nucleoside and, hence, provides a spectrophotometric signal that can be continuously followed. In the presence of excess phosphatase, the phosphorylated protein substrate molecules undergo dephosphorylation almost immediately after their formation. The steady-state use of the resultant inorganic phosphate is a reflection of the constant initial velocity of the exchange reaction. Method development and evaluation, overview | Homo sapiens | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
MAPK kinase | - |
Homo sapiens |
MAPK kinase 6 | - |
Homo sapiens |
MAPK/ERK kinase 1 | - |
Homo sapiens |
MEK1 | - |
Homo sapiens |
MKK6 | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.074 | - |
K52R-[ERK2] | pH 7.0, 25°C | Homo sapiens | |
0.085 | - |
ATP | pH 7.0, 25°C | Homo sapiens | |
0.152 | - |
K53M-[p38alpha] | pH 7.0, 25°C | Homo sapiens | |
0.243 | - |
p38alpha | pH 7.0, 25°C | Homo sapiens | |
0.503 | - |
ERK2 | pH 7.0, 25°C | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Homo sapiens |
General Information | Comment | Organism |
---|---|---|
metabolism | MEK1 activates the prototypic MAPK pathway by activating enzyme ERK1 | Homo sapiens |
metabolism | the p38 pathway is activated by MKK3, MKK4, and MKK6 | Homo sapiens |