DGKE Kinase Enzyme System

Recombinant full-length human DGKE was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. Diacylglycerol kinase epsilon (DAG kinase epsilon) is the only member of class III diacylglycerol kinase family, which lacks the E-F hand motifs for Ca2+ binding but possess a clear selectivity for arachidonoyl-containing species of diacylglycerol. DGK epsilon might decrease signals conducted via arachidonoyl-diacylglycerol or promotes the synthesis of phospholipids with certain fatty acids (1). DGK epsilon contributes to Huntington disease pathogenesis and its inhibition might be effect in the therapeutics. ADP-Glo Kinase Assay is a luminescent kinase assay that measures ADP formed from a kinase reaction; ADP is converted into ATP, which is a substrate in a reaction catalyzed by Ultra-Glo Luciferase that produces light. The luminescent signal positively correlates with ADP amount and kinase activity. The assay is well suited for measuring the effects chemical compounds have on the activity of a broad range of purified kinases, making it ideal for both primary screening as well as kinase selectivity profiling. The ADP-Glo Kinase Assay can be used to monitor the activity of virtually any ADP-generating enzyme (e.g., kinase or ATPase) using up to 1mM ATP.