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Literature summary extracted from

  • Kenski, D.M.; Zhang, C.; von Zastrow, M.; Shokat, K.M.
    Chemical genetic engineering of G protein-coupled receptor kinase 2 (2005), J. Biol. Chem., 280, 35051-35061.
    View publication on PubMed

Application

EC Number Application Comment Organism
2.7.11.15 biotechnology engineering of a GRK2 mutant sensitive to a specific inhibitor Bos taurus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.11.15 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

EC Number Protein Variants Comment Organism
2.7.11.15 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
2.7.11.15 L271G site-directed mutagenesis, mutant GRK2 shows highly reduced activity compared to the wild-type enzyme Bos taurus
2.7.11.15 L273Y site-directed mutagenesis, mutant GRK2 shows highly reduced activity compared to the wild-type enzyme Bos taurus
2.7.11.15 L336F site-directed mutagenesis, mutant GRK2 shows highly reduced activity compared to the wild-type enzyme Bos taurus

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.11.15 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.11.15 0.017
-
ATP pH 7.8, 23°C, recombinant wild-type GRK2, with substrate tubulin Bos taurus
2.7.11.15 0.062
-
ATP pH 7.8, 23°C, recombinant GRK2 L271G mutant, with substrate tubulin Bos taurus
2.7.11.15 0.149
-
ATP pH 7.8, 23°C, recombinant GRK2 C221V mutant, with substrate tubulin Bos taurus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.11.15 Mg2+
-
Bos taurus

Organism

EC Number Organism UniProt Comment Textmining
2.7.11.15 Bos taurus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.11.15 recombinant His-tagged wild-type and mutant GRK2s from Spodoptera frugiperda Sf9 cells by nickel affinity chromatography Bos taurus

Storage Stability

EC Number Storage Stability Organism
2.7.11.15 -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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.11.15 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
-
?
2.7.11.15 ATP + rhodopsin
-
Bos taurus ADP + phosphorylated rhodopsin
-
?
2.7.11.15 ATP + tubulin
-
Bos taurus ADP + phosphorylated tubulin
-
?

Synonyms

EC Number Synonyms Comment Organism
2.7.11.15 G protein-coupled receptor kinase 2
-
Bos taurus
2.7.11.15 GRK2
-
Bos taurus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.11.15 23
-
assay at Bos taurus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.11.15 0.0001
-
ATP pH 7.8, 23°C, recombinant GRK2 L271G mutant, with substrate tubulin Bos taurus
2.7.11.15 0.0029
-
ATP pH 7.8, 23°C, recombinant wild-type GRK2, with substrate tubulin Bos taurus
2.7.11.15 0.0041
-
ATP pH 7.8, 23°C, recombinant GRK2 C221V mutant, with substrate tubulin Bos taurus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.11.15 7.8
-
assay at Bos taurus

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.11.15 ATP
-
Bos taurus