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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 PHKG1 with 0.25 mg/ml ZIKtide peptide 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: PHKG1 or phosphorylase kinase gamma 1 is a member of the Ser/Thr protein kinase family that encodes a protein with one protein kinase domain and two calmodulin-binding domains. Phosphorylase kinase is a crucial glycogenolytic regulatory enzyme (1). PHKG1 is the catalytic member of a 16-subunit protein kinase complex that contains equimolar ratios of 4 subunit types known as alpha, beta, gamma and delta (2). Skeletal muscle contains the highest amount of phosphorylase kinase enzymatic activity, although activity is also observed in liver, cardiac muscle, brain, and several other tissues.
Description: Recombinant human PHKG1 (phosphorylase kinase gamma 1), GenBank Accession No. NM_006213, full length, with N-terminal GST-tag, expressed in Sf9 insect cells via a baculovirus expression system.
Format: Aqueous buffer solution
Formulation: 50 mM Tris-HCl, pH 7.5, 50 mM NaCl, 0.25 mM DTT, 0.1 mM EDTA, 0.1 mM PMSF, 25% glycerol.
Genbank: NM_006213
Host Cell Line: Sf9 cells
Purity: ≥75%
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: Q16816
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Burwinkel, B. et.al: Muscle glycogenosis with low phosphorylase kinase activity: mutations in PHKA1, PHKG1 or six other candidate genes explain only a minority of cases. Europ. J. Hum. Gen. 11: 516-526, 2003.
2. Wehner, M. et.al: Human cDNA encoding the muscle isoform of the phosphorylase kinase gamma subunit (PHKG1).
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