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Literature summary extracted from

  • Lodowski, D.T.; Barnhill, J.F.; Pyskadlo, R.M.; Ghirlando, R.; Sterne-Marr, R.; Tesmer, J.J.
    The role of Gbetagamma and domain interfaces in the activation of G protein-coupled receptor kinase 2 (2005), Biochemistry, 44, 6958-6970.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.7.11.15 Gbetagamma subunits required for activity on G protein-coupled receptors, binding via enzyme C-terminal pleckstrin homology domain, binding does not induce large domain rearrangements, but small rotations of the enzyme domains, the domain interfaces remain intact upon GRK2 activation, structure analysis of GRK2 in complex with G protein beta1gamma2 subunits, overview Bos taurus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.11.15 co-overexpression of GRK2 mutants and G protein beta1gamma2 subunits in Spodoptera frugiperda Sf9 cells using the baculovirus infection system, and in COS-1 cells Bos taurus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.11.15 purified recombinant GRK2 S670A mutant, hanging drop vapour diffusion method, 4°C, 0.002 ml protein solution containing 15 mg/ml GRK2, 20 mM HEPES, pH 8.0, 0.2 M NaCl, and 2 mM DTT, mixed with 0.002 ml well solution containing 0.1 M HEPES, pH 7.5-8.0, 1 M NaCl, 1 M urea, 10-40 mM phosphoserine, pH 7.0, 5% v/v glycerol, and 9.5-11.5% PEG 8000, 2-3 weeks, X-ray diffraction structure determination and analysis at 4.5 A resolution Bos taurus

Protein Variants

EC Number Protein Variants Comment Organism
2.7.11.15 E520A site-directed mutagenesis, very low recombinant expression level, mutant GRK2 shows a dramatic loss of activity with rhodopsin as substrate Bos taurus
2.7.11.15 E96A site-directed mutagenesis, slightly increased recombinant expression level compared to the wild-type GRK2, mutant GRK2 shows activity with rhodopsin as substrate similar to the wild-type enzyme Bos taurus
2.7.11.15 P638D site-directed mutagenesis, RH-PH domain interface residue mutant, 40% reduced recombinant expression level compared to the wild-type GRK2, mutant GRK2 shows 15% of wild-type activity with rhodopsin as substrate Bos taurus
2.7.11.15 R516A site-directed mutagenesis, 60% reduced recombinant expression level compared to the wild-type GRK2, mutant GRK2 shows slightly reduced activity with rhodopsin as substrate Bos taurus
2.7.11.15 S670A site-directed mutagenesis, the MAP kinase phosphorylation site S670 is eliminated in the GRK2 mutant enzyme Bos taurus
2.7.11.15 V42E site-directed mutagenesis, RH-PH domain interface residue mutant, 60% reduced recombinant expression level compared to the wild-type GRK2, mutant GRK2 shows 25% of wild-type activity with rhodopsin as substrate Bos taurus
2.7.11.15 Y46A site-directed mutagenesis, RH-PH domain interface residue mutant, 80% reduced recombinant expression level compared to the wild-type GRK2, mutant GRK2 shows 50% of wild-type activity with rhodopsin as substrate Bos taurus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.7.11.15 membrane associated, GRK2 in complex with G protein beta1gamma2 subunits Bos taurus 16020
-

Metals/Ions

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

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.11.15 additional information
-
 equilibrium sedimentation measurements on GRK2-Gbetagamma complexes, overview Bos taurus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.11.15 ATP + G protein-coupled receptor Bos taurus GRK2 performs desensitization of the ligand-activated receptor by phosphorylation ADP + phosphorylated G protein-coupled receptor
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.11.15 Bos taurus P21146
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
2.7.11.15 phosphoprotein GRK2 is phosphorylated by MAP kinase at S670 Bos taurus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.11.15 ATP + G protein-coupled receptor
-
 Bos taurus ADP + phosphorylated G protein-coupled receptor
-
 ?
2.7.11.15 ATP + G protein-coupled receptor GRK2 performs desensitization of the ligand-activated receptor by phosphorylation Bos taurus ADP + phosphorylated G protein-coupled receptor
-
 ?
2.7.11.15 ATP + rhodopsin dark-adapted and light-activated rhodopsin in urea-washed rod outer segment membranes Bos taurus ADP + phosphorylated rhodopsin
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.11.15 More GRK2 consists of 3 modular domains: a predominantly N-terminal RGS homology domain, a central protein kinase domain, and a C-terminal pleckstrin homology domain, the interfaces of the domains are important for stability, and enzyme expression and activity, and remain intact upon GRK2 activation, structure analysis of GRK2 in complex with G protein beta1gamma2 subunits, overview Bos taurus

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 30
-
 assay at Bos taurus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.11.15 7.5
-
 assay at Bos taurus

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.11.15 ATP
-
 Bos taurus