Binding affinity to human recombinant UP1-R1P binary complex expressed in Escherichia coli Rosetta (DE3) at 200 uM R1P by isothermal titration calorimetric analysis
Binding affinity to human recombinant UP1-R1P binary complex expressed in Escherichia coli Rosetta (DE3) at 150 uM R1P by isothermal titration calorimetric analysis
Binding affinity to human recombinant UP1-R1P binary complex expressed in Escherichia coli Rosetta (DE3) at 50 uM R1P by isothermal titration calorimetric analysis
Binding affinity to human recombinant UP1-inorganic phosphate binary complex expressed in Escherichia coli Rosetta (DE3) by isothermal titration calorimetric analysis
Uncompetitive inhibition of human recombinant UP1 expressed in Escherichia coli Rosetta (DE3) using inorganic phosphate as substrate by Lineweaver-Burk plot analysis
Non-competitive inhibition of human recombinant UP1 expressed in Escherichia coli Rosetta (DE3) using inorganic phosphate as substrate assessed as equilibrium dissociation constant for enzyme-substrate-inhibitor complex by Lineweaver-Burk plot analysis
Non-competitive inhibition of human recombinant UP1 expressed in Escherichia coli Rosetta (DE3) using inorganic phosphate as substrate assessed as equilibrium dissociation constant for enzyme-inhibitor complex by Lineweaver-Burk plot analysis
Non-competitive inhibition of human recombinant UP1 expressed in Escherichia coli Rosetta (DE3) using URD as substrate assessed as equilibrium dissociation constant for enzyme-substrate-inhibitor complex by Lineweaver-Burk plot analysis
Time dependent inhibition of human recombinant UP1 expressed in Escherichia coli Rosetta (DE3) using URD and inorganic phosphate as substrate at 1 uM by spectrophotometric analysis
Inhibition of human recombinant UP1 expressed in Escherichia coli Rosetta (DE3) using URD and inorganic phosphate as substrate at 1 uM after 60 seconds by spectrophotometric analysis relative to control
Non-competitive inhibition of human recombinant UP1 expressed in Escherichia coli Rosetta (DE3) using URD as substrate assessed as equilibrium dissociation constant for enzyme-inhibitor complex by Lineweaver-Burk plot analysis
Enzyme Assay: The aim of this study was to evaluate TK-112690 in vivo as an inhibitor of uridine phosphorylase (UPase) enzyme activity. The range of TK-112690 doses studied for ability to prevent metabolic breakdown of uridine, through the in vitro inhibition of mouse and human small intestinal UPase enzyme, was 0, 0.1, 0.5, 1, 5, 10, 50, 100, 500, 1000, 5000 and 10000 uM). Detection of UPase activity was determined by HPLC analysis using UV detection of uracil concentration (UPase catabolizes uridine into uracil and ribose-1-phosphate).The UPase enzyme material was prepared from homogenized mouse and human being small intestinal tissue. TK-112690 was dissolved in water (50 mg/ml) and analyzed for UPase inhibition in aqueous solution containing 5 mM uridine, 0.01 M Tris, 0.01 M phosphate, 1 mM EDTA, and 1 mM DTT. Reactions were performed at 37 C. at pH of 7.3.TK-11260 inhibition of mouse and human UPase was analyzed by reverse phase HPLC using UV detection.