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Literature summary for 2.7.10.2 extracted from

  • Lin, X.; Lee, S.; Sun, G.
    Functions of the activation loop in Csk protein-tyrosine kinase (2003), J. Biol. Chem., 278, 24072-24077.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
additional information Csk is tyrosine autophosphorylated in the activation loop, structure Homo sapiens

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant His-tagged Csk in Escherichia coli strain BL21(DE3) Homo sapiens

Protein Variants

Protein Variants Comment Organism
D332A site-directed mutagenesis of Csk in the DFG motif in front of the activation loop leads to complete loss of kinase activity Homo sapiens
D344A site-directed mutagenesis of Csk in the activation loop, mutant shows slightly increased activity with kdSrc and slightly reduced activity with poly(Glu4-Tyr) as substrates compared to the wild-type enzyme Homo sapiens
E338A site-directed mutagenesis of Csk in the activation loop, mutant shows 40-60% remaining activity compared to the wild-type enzyme Homo sapiens
G346A site-directed mutagenesis of Csk in the activation loop, mutant shows decreased activity compared to the wild-type enzyme Homo sapiens
K337A site-directed mutagenesis of Csk in the activation loop, mutant shows slightly decreased activity compared to the wild-type enzyme Homo sapiens
additional information construction of a residue-339-deletion mutant Homo sapiens
Q343A site-directed mutagenesis of Csk in the activation loop, mutant shows slightly decreased activity compared to the wild-type enzyme Homo sapiens
R313A site-directed mutagenesis, activity similar to the wild-type enzyme Homo sapiens
S340A site-directed mutagenesis of Csk in the activation loop, mutant shows slightly decreased activity compared to the wild-type enzyme Homo sapiens
S341A site-directed mutagenesis of Csk in the activation loop, mutant shows slightly decreased activity compared to the wild-type enzyme Homo sapiens
T336A site-directed mutagenesis of Csk in the activation loop, mutant shows increased activity compared to the wild-type enzyme Homo sapiens
T342A site-directed mutagenesis of Csk in the activation loop, mutant shows decreased activity compared to the wild-type enzyme Homo sapiens
T345A site-directed mutagenesis of Csk in the activation loop, mutant shows decreased activity compared to the wild-type enzyme Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.01
-
[kdSrc kinase]-L-tyrosine pH 8.0, 30°C, recombinant mutant E338A Csk Homo sapiens
0.01
-
[kdSrc kinase]-L-tyrosine pH 8.0, 30°C, recombinant wild-type Csk Homo sapiens
0.24
-
[kdSrc kinase]-L-tyrosine pH 8.0, 30°C, recombinant Csk residue-339-deletion mutant Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
phosphoprotein Csk is tyrosine autophosphorylated in the activation loop Homo sapiens

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant His-tagged Csk from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + poly(Glu4-Tyr)
-
Homo sapiens ADP + poly(Glu4-Tyr)-L-tyrosine phosphate
-
?
ATP + [kdSrc kinase]-L-tyrosine i.e. kinase-defective chicken Src mutant K295M Homo sapiens ADP + [kdSrc kinase]-L-tyrosine phosphate
-
?
additional information the activation loop is not essential for enzyme activity Homo sapiens ?
-
?

Synonyms

Synonyms Comment Organism
Csk protein-tyrosine kinase
-
Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.01
-
poly(Glu4-Tyr) pH 8.0, 30°C, recombinant Csk residue-339-deletion mutant Homo sapiens
0.1
-
poly(Glu4-Tyr) pH 8.0, 30°C, recombinant mutant E338A Csk Homo sapiens
0.35
-
[kdSrc kinase]-L-tyrosine pH 8.0, 30°C, recombinant Csk residue-339-deletion mutant Homo sapiens
0.57
-
poly(Glu4-Tyr) pH 8.0, 30°C, recombinant wild-type Csk Homo sapiens
0.82
-
[kdSrc kinase]-L-tyrosine pH 8.0, 30°C, recombinant mutant E338A Csk Homo sapiens
1.88
-
[kdSrc kinase]-L-tyrosine pH 8.0, 30°C, recombinant wild-type Csk Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Homo sapiens

Cofactor

Cofactor Comment Organism Structure
ATP
-
Homo sapiens