ADP-Glo Kinase Assay + PRKG1 (PKG) Kinase Enzyme System
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. Kinase Enzyme System contains: Kinase: PDK1, 10ug (Human, recombinant full-length). MW: ~67kDa. Substrate: PDKtide (KTFCGTPEYLAPEVRREPRILSEEEQEMFRDFDYIADWC); derived from two human proteins: residues 1-14 are based on AKT1 (307-320) and residues 16-39 are based on PKN2/PRK2 (961-984). Other: Reaction Buffer, DTT. PDK1 NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/5170/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.