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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 GRK5 with 0.5 mg/ml casein 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: GRK5 is a member of the guanine nucleotide-binding protein (G protein)-coupled receptor kinase subfamily of the Ser/Thr protein kinase family (1). It phosphorylates the activated forms of G protein-coupled receptors thus initiating their deactivation. GRK5 plays a role in regulating the motility of polymorphonuclear leukocytes (PMNs). Desensitization of G protein-coupled receptors regulates the number of polymorphonuclear leukocytes (PMNs), as well as their motility and ability to stop upon contact with pathogens or target cells, and this desensitization is mediated by GRK5 (2).
Description: Human GRK5, also known as GPRK5 (GenBank Accession No. NM_005308), full length, with N-terminal GST tag, MW ~95 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, 0.25 mM DTT, 0.1 mM EGTA, 0.1 mM EDTA, 0.1 mM PMSF, 25% glycerol.
Genbank: NM_005308
Host Cell Line: Sf9 cells
Purity: ≥90%
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: P34947
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Haribabu, B. et al: Identification of additional members of human G-protein-coupled receptor kinase multigene family. Proc. Nat. Acad. Sci. 90: 9398-9402, 1993.
2. Fan, J. et al: Toll-like receptor-4 (TLR4) signaling augments chemokine-induced neutrophil migration by modulating cell surface expression of chemokine receptors. Nature Med. 9: 315-321, 2003.
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