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Literature summary for 2.7.12.2 extracted from

  • Zheng, L.S.; Zhang, Y.Y.; Wu, J.W.; Wu, Z.; Zhang, Z.Y.; Wang, Z.X.
    A continuous spectrophotometric assay for mitogen-activated protein kinase kinases (2012), Anal. Biochem., 421, 191-197.
    View publication on PubMed
Search for more literature for 2.7.12.2

Application

Application Comment Organism
analysis development of a continuous spectrophotometric assay for mitogen-activated protein kinase kinases Homo sapiens

Protein Variants

Protein Variants Comment Organism
S207E/T211E a constitutively active MKK6 mutant Homo sapiens

KM Value [mM]

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

Metals/Ions Comment Organism Structure
Mg2+ required Homo sapiens

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Homo sapiens P52564
-
-
Homo sapiens Q02750
-
-

Substrates and Products (Substrate)

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

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Homo sapiens

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 assay at Homo sapiens

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Homo sapiens

General Information

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