Literature summary for 2.7.11.15 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.

Search for more literature for 2.7.11.15

Activating Compound

Activating Compound	Comment	Organism	Structure
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)

Cloned (Comment)	Organism
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)

Crystallization (Comment)	Organism
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

Crystallization (Comment)

Organism

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

Protein Variants	Comment	Organism
E520A	site-directed mutagenesis, very low recombinant expression level, mutant GRK2 shows a dramatic loss of activity with rhodopsin as substrate	Bos taurus
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
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
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
S670A	site-directed mutagenesis, the MAP kinase phosphorylation site S670 is eliminated in the GRK2 mutant enzyme	Bos taurus
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
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

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

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	-	Bos taurus

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Bos taurus	P21146	-	-

Posttranslational Modification

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

Substrates and Products (Substrate)

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

Subunits

Subunits	Comment	Organism
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

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
30	-	assay at	Bos taurus

pH Optimum

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

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Bos taurus