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Literature summary for 2.7.11.15 extracted from
- Chemical genetic engineering of G protein-coupled receptor kinase 2 (2005), J. Biol. Chem., 280, 35051-35061.
Application
| Application | Comment | Organism |
|---|---|---|
| biotechnology | engineering of a GRK2 mutant sensitive to a specific inhibitor | Bos taurus |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| stable co-expression of C-terminally HA-tagged wild-type GRK2 and murine micro-opioid receptor in HEK-293 cells, transient expression of C-terminally HA-tagged wild-type and mutant GRK2s in HEK-293 cells, expression of His-tagged wild-type and mutant GRK2s in Spodoptera frugiperda Sf9 cells using the baculovirus infection system | Bos taurus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| C221V | site-directed mutagenesis, mutant GRK2 activity is slightly increased compared to the wild-type enzyme, the requirement for initial ligand-induced internalization of a G protein-coupled receptor with subsequent rounds of internalization is different for the mutant GRK2 compared to the wild-type enzyme | Bos taurus |
| L271G | site-directed mutagenesis, mutant GRK2 shows highly reduced activity compared to the wild-type enzyme | Bos taurus |
| L273Y | site-directed mutagenesis, mutant GRK2 shows highly reduced activity compared to the wild-type enzyme | Bos taurus |
| L336F | site-directed mutagenesis, mutant GRK2 shows highly reduced activity compared to the wild-type enzyme | Bos taurus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 4-amino-1-tert-butyl-3-(1'-naphthyl)pyrrazolo[3,4-d]pyrimidine | i.e. 1-Na-PP1, inhibits mutant C221V mediated morphine-induced internalization of the micro-opioid receptor, but not wild-type GRK2 and mutant L271G activities | Bos taurus |  |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.017 | - |
ATP | pH 7.8, 23°C, recombinant wild-type GRK2, with substrate tubulin | Bos taurus | |
| 0.062 | - |
ATP | pH 7.8, 23°C, recombinant GRK2 L271G mutant, with substrate tubulin | Bos taurus | |
| 0.149 | - |
ATP | pH 7.8, 23°C, recombinant GRK2 C221V mutant, with substrate tubulin | Bos taurus | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | - |
Bos taurus | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bos taurus | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged wild-type and mutant GRK2s from Spodoptera frugiperda Sf9 cells by nickel affinity chromatography | Bos taurus |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -80°C, purified recombinant His-tagged wild-type and mutant GRK2s, 50 mM NaH2PO4, pH 8.0, 300 mM NaCl, 250 mM imidazole, 30% glycerol, stable for 2 months | Bos taurus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + micro-opioid receptor | mutant C221V GRK2 shows slightly higher effect on morphine-induced internalization of the micro-opioid receptor compared to the wild-type GRK2, while the effects of mutants L271G, L273Y, and L336F are reduced, overview | Bos taurus | ADP + phosphorylated micro-opioid receptor | - |
? | |
| ATP + rhodopsin | - |
Bos taurus | ADP + phosphorylated rhodopsin | - |
? | |
| ATP + tubulin | - |
Bos taurus | ADP + phosphorylated tubulin | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| G protein-coupled receptor kinase 2 | - |
Bos taurus |
| GRK2 | - |
Bos taurus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 23 | - |
assay at | Bos taurus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0001 | - |
ATP | pH 7.8, 23°C, recombinant GRK2 L271G mutant, with substrate tubulin | Bos taurus | |
| 0.0029 | - |
ATP | pH 7.8, 23°C, recombinant wild-type GRK2, with substrate tubulin | Bos taurus | |
| 0.0041 | - |
ATP | pH 7.8, 23°C, recombinant GRK2 C221V mutant, with substrate tubulin | Bos taurus | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.8 | - |
assay at | Bos taurus |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| ATP | - |
Bos taurus | |
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