Recombinant human WNK1 (181–507) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. WNK1 is a member of the WNK subfamily of serine/threonine protein kinases that is a key regulator of blood pressure by controlling the transport of sodium and chloride ions. Mutations in WNK1 have been associated with pseudohypoaldosteronism type II and hereditary sensory neuropathy type II. WNK1 is a regulator of blood pressure and deficiency of this protein in mice lowers the blood pressure (1). WNK1 can regulate the Ca2+-sensing and the subsequent Ca2+-dependent interactions mediated by synaptotagmin C2 domains, and WNK1 exhibits additive CFTR inhibition (2).
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.
Profile More Compounds In-House: ADP-Glo™ Kinase Assay + Kinase Enzyme System is optimized so that you are up and running in no time. Complete Systems: The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer, DTT and supplemental reagents as needed. Obtain Reliable Results: The broad dynamic range, the ease of use and better sensitivity obtained with ADP-Glo™ Kinase Assay result in less ambiguous data.
Notes
Kinase Enzyme System manufactured by SignalChem.
Bulk quantities available upon request.
References
1.Zambrowicz, B.P. et al. (2003) Wnk1 kinase deficiency lowers blood pressure in mice: a gene-trap screen to identify potential targets for therapeutic intervention. Proc. Natl. Acad. Sci. USA 100, 14109–114.
2.Lee, B.-H. et al. (2004) WNK1 phosphorylates synaptotagmin 2 and modulates its membrane binding. Molec. Cell 15, 741–51.