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Literature summary for 2.7.10.2 extracted from

  • Kemble, D.J.; Wang, Y.H.; Sun, G.
    Bacterial expression and characterization of catalytic loop mutants of SRC protein tyrosine kinase (2006), Biochemistry, 45, 14749-14754.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
additional information the enzyme is activated by phosphorylation, and also performs autophosphorylation Homo sapiens

Application

Application Comment Organism
drug development tyrosine kinase Src is a key enzyme in mammalian signal transduction and an important target for anticancer drug discovery Homo sapiens

Cloned(Commentary)

Cloned (Comment) Organism
expression of active Src in Escherichia coli strain BL21(DE3) is difficult and ineffective due to toxicity caused by Src kinase activity, effective expression of inactive mutants, e.g. of K295M, is possible, overview Homo sapiens

Protein Variants

Protein Variants Comment Organism
H384A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type Src Homo sapiens
H384Q site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type Src Homo sapiens
H384Q/Y416F site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type Src Homo sapiens
H384Y site-directed mutagenesis, the mutant shows reduced activity and no autophosphorylation compared to the wild-type Src Homo sapiens
H384Y/Y416F site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type Src Homo sapiens
H384Y/Y527F site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type Src Homo sapiens
K295M site-directed mutagenesis, inactive Src mutant Homo sapiens
N391D site-directed mutagenesis, the mutant shows highly reduced enzyme activity and can be well expressed and purified using the Escherichia coli expression system Homo sapiens
R388A/A390R site-directed mutagenesis, mutations in the catalytic loop, the mutant shows slightly reduced enzyme activity and cannot be well expressed and purified using the Escherichia coli expression system Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information
-
Homo sapiens
0.075
-
ATP pH 8.0, 30°C, recombinant wild-type enzyme Homo sapiens
0.253
-
ATP pH 8.0, 30°C, recombinant mutant H384Y/Y527F Homo sapiens
0.309
-
ATP pH 8.0, 30°C, recombinant mutant H384Y Homo sapiens
0.327
-
ATP pH 8.0, 30°C, recombinant mutant H384Q Homo sapiens
0.335
-
ATP pH 8.0, 30°C, recombinant mutant H384Y/Y416F Homo sapiens
0.34
-
ATP pH 8.0, 30°C, recombinant mutant H384A Homo sapiens
0.345
-
ATP pH 8.0, 30°C, recombinant mutant H384Q/Y416F Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Homo sapiens tyrosine kinase Src is a key enzyme in mammalian signal transduction and an important target for anticancer drug discovery ?
-
?

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
phosphoprotein the enzyme is activated by phosphorylation, and also performs autophosphorylation Homo sapiens

Purification (Commentary)

Purification (Comment) Organism
recombinant inactive Src mutants from Escherichia coli strain BL21(DE3) by affinity chromatography, Src mutants with kinase activity above a certain threshold cannot be purified from a bacterial expression system, while a variety of mutants with a kinase activity below this threshold can be expressed and purified, overview Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + EEEEY
-
Homo sapiens ADP + EEEE(phospho)Y
-
?
additional information tyrosine kinase Src is a key enzyme in mammalian signal transduction and an important target for anticancer drug discovery Homo sapiens ?
-
?
additional information the enzyme is activated by phosphorylation, and also performs autophosphorylation Homo sapiens ?
-
?

Synonyms

Synonyms Comment Organism
Src
-
Homo sapiens
Src 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
0.072
-
EEEEY pH 8.0, 30°C, recombinant mutant H384Y/Y416F Homo sapiens
0.1
-
EEEEY pH 8.0, 30°C, recombinant mutant H384A Homo sapiens
0.15
-
EEEEY pH 8.0, 30°C, recombinant mutant H384Q/Y416F Homo sapiens
0.16
-
ATP pH 8.0, 30°C, recombinant mutant H384Y/Y416F Homo sapiens
0.21
-
ATP pH 8.0, 30°C, recombinant mutant H384A Homo sapiens
0.22
-
EEEEY pH 8.0, 30°C, recombinant mutant H384Y Homo sapiens
0.4
-
ATP pH 8.0, 30°C, recombinant mutant H384Q/Y416F Homo sapiens
0.43
-
EEEEY pH 8.0, 30°C, recombinant mutant H384Y/Y527F Homo sapiens
0.53
-
ATP pH 8.0, 30°C, recombinant mutant H384Y Homo sapiens
0.8
-
EEEEY pH 8.0, 30°C, recombinant mutant H384Q Homo sapiens
0.98
-
ATP pH 8.0, 30°C, recombinant mutant H384Y/Y527F Homo sapiens
1.93
-
ATP pH 8.0, 30°C, recombinant mutant H384Q Homo sapiens
2.27
-
EEEEY pH 8.0, 30°C, recombinant wild-type enzyme Homo sapiens
3.13
-
ATP pH 8.0, 30°C, recombinant wild-type enzyme Homo sapiens
3.52
-
ATP pH 8.0, 30°C, recombinant wild-type enzyme 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