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Literature summary extracted from

  • Singh, P.; Wang, B.; Maeda, T.; Palczewski, K.; Tesmer, J.J.
    Structures of rhodopsin kinase in different ligand states reveal key elements involved in G protein-coupled receptor kinase activation (2008), J. Biol. Chem., 283, 14053-14062.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.11.14 expression of His6-tagged wild-type and mutant enzymes in High Five insect cells using the baculovirus transfection system Bos taurus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.11.14 purified recombinant enzyme, X-ray diffraction structure determination and analysis, molecular replacement Bos taurus

Protein Variants

EC Number Protein Variants Comment Organism
2.7.11.14 D164A site-directed mutagenesis, a dimer interface mutant showing only slightly altered kinetics compared to the wild-type enzyme Bos taurus
2.7.11.14 D164A/L166K site-directed mutagenesis, a dimer interface mutant showing reduced activity compared to the wild-type enzyme Bos taurus
2.7.11.14 D164A/W531A site-directed mutagenesis, inactive mutant Bos taurus
2.7.11.14 L166K site-directed mutagenesis, a dimer interface mutant showing reduced activity compared to the wild-type enzyme Bos taurus
2.7.11.14 L166K/W531A site-directed mutagenesis, inactive mutant Bos taurus
2.7.11.14 S5A site-directed mutagenesis, an N-terminal mutant showing reduced activity compared to the wild-type enzyme Bos taurus
2.7.11.14 S5D site-directed mutagenesis, inactive mutant Bos taurus
2.7.11.14 T8A site-directed mutagenesis, an N-terminal mutant showing reduced activity compared to the wild-type enzyme Bos taurus
2.7.11.14 T8D site-directed mutagenesis, inactive mutant Bos taurus
2.7.11.14 T8E site-directed mutagenesis, an N-terminal mutant showing reduced activity compared to the wild-type enzyme Bos taurus
2.7.11.14 W531A site-directed mutagenesis, inactive mutant Bos taurus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.11.14 additional information
-
 additional information steady-state kinetics Bos taurus
2.7.11.14 0.0015
-
 rhodopsin pH 7.5, 30°C, mutant T8A Bos taurus
2.7.11.14 0.0021
-
 rhodopsin pH 7.5, 30°C, wild-type enzyme Bos taurus
2.7.11.14 0.0031
-
 rhodopsin pH 7.5, 30°C, mutant D164A/L166K Bos taurus
2.7.11.14 0.0038
-
 rhodopsin pH 7.5, 30°C, mutant D164A Bos taurus
2.7.11.14 0.0048
-
 rhodopsin pH 7.5, 30°C, mutant T8E Bos taurus
2.7.11.14 0.0051
-
 rhodopsin pH 7.5, 30°C, mutant S5A Bos taurus
2.7.11.14 0.0067
-
 rhodopsin pH 7.5, 30°C, mutant L166K Bos taurus

Metals/Ions

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

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.11.14 ATP + rhodopsin Bos taurus
-
 ADP + phosphorhodopsin
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.11.14 Bos taurus P28327
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.11.14 recombinant His-tagged wild-type and mutant enzymes from High Five insect cells by nickel affinity chromatography and ion exchange chromatography Bos taurus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.11.14 ATP + rhodopsin
-
 Bos taurus ADP + phosphorhodopsin
-
 ?
2.7.11.14 additional information nucleotide binding and conformational change, overview Bos taurus ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.11.14 More domain structure analysis, mass spectrometric analysis, overview Bos taurus

Synonyms

EC Number Synonyms Comment Organism
2.7.11.14 GRK1
-
 Bos taurus
2.7.11.14 More GRK1 is the founding member of the G protein-coupled receptor kinase, GRK kinase, family Bos taurus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.11.14 30
-
 assay at Bos taurus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.11.14 7.5
-
 assay at Bos taurus

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.11.14 ATP
-
 Bos taurus