Recombinant full-length human PRKG1 (PKG) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. PRKG1 is a homodimer, with each monomer containing a regulatory cGMP-binding domain and a catalytic domain (1). PRKG1 was shown to be expressed at highest levels in bladder, uterus, adrenal gland and fallopian tube. PRKG1 plays an important stimulatory role in platelet activation (2). Expression of recombinant PRKG1 in a reconstituted cell model enhanced von Willebrand factor-induced activation of the platelet integrin alpha-IIb/beta-3. PRKG1 knockout mice showed impaired platelet responses to VWF or low doses of thrombin and prolonged bleeding time. Human platelet aggregation induced by VWF or low-dose thrombin was inhibited by PRKG1 inhibitors but enhanced by cGMP.
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
Orstavik, S.et al.(1997) Characterization of the human gene encoding the type I-alpha and type I-beta cGMP-dependent protein kinase (PRKG1).Genomics42, 311–318.
Li, Z.et al.(2003) A stimulatory role for cGMP-dependent protein kinase in platelet activation.Cell112, 77–86.