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Encompassing Amino Acids: full length
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: 5 mM MOPS, 2.5 mM β-glycerophosphate, 5 mM MgCl2, 1 mM EGTA, 0.4 mM EDTA, 0.05 mM DTT, 2 mM ATP. Incubate GRK7 with 0.25 mg/ml Myelin basic Protein (substrate) and 50 ?M [33P]-ATP at 30°C for 15 minutes, then spot reaction on phosphocellulose paper, fix in 1% phosphoric acid, and assay with a scintillation counter.
Background: GRK7 is a member of the G protein-coupled receptor kinase subfamily and is a Ser/Thr protein kinase. GRK7 is specifically expressed in the retina and has been shown to phosphorylate cone opsins and initiate their deactivation (1). GRK7 catalyzes rhodopsin phosphorylation in a light-dependent manner. GRK7 co-localizes with GRK1 in human cone outer segments. PKA phosphorylates GRK7 at Ser(23) and Ser(36) in vitro and phosphorylation of GRK7 by PKA reduces the ability of GRK7 to phosphorylate rhodopsin in vitro (2). Since exposure to light causes a decrease in cAMP levels in rod cells, it is proposed that phosphorylation of GRK7 by PKA occurs in the dark.
Description: Human GRK7, also known as GPRK7 (GenBank Accession No. NM_139209), full length, with N-terminal GST tag, MW ~89 kDa, expressed in Sf9 insect cells via a baculovirus expression system.
Format: Aqueous buffer solution
Formulation: 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10 mM glutathione, 0.1 mM EDTA, 0.25 mM DTT, 0.1 mM PMSF, 25% glycerol
Genbank: NM_139209
Host Cell Line: Sf9 cells
Purity: ≥90%
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: Q8WTQ7
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Chen, C-K. et al: Characterization of human GRK7 as a potential cone opsin kinase. Molec. Vision 7: 305-313, 2001.
2. Horner, T J. et al: Phosphorylation of GRK1 and GRK7 by cAMP-dependent protein kinase attenuates their enzymatic activities. J Biol Chem. 2005 Aug 5;280(31):28241-50.
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