Patent ID: 6723499
Filing Date: 2004-04-20
Classification: C12Q

Abstract:
A method for identifying an inhibitor of a dual substrate enzyme; wherein a first substrate is a macromolecule that is enzymatically modified, in the presence of the dual substrate enzyme, to accept a radiolabeled portion of a second substrate, said method comprising:a. adding a capture resin to a buffered mixture of an enzyme, allowing the enzyme to catalyze transfer of the radiolabeled portion of the radiolabeled second substrate to a non-radiolabeled first substrate, in the presence or absence of a test compound; b. removing unreacted radiolabeled second substrate; c. adding a scintillant resin to an enzyme-radiolabeled first substrate mixture; d. measuring the amount of radiolabeled first substrate reacted in the presence of the test compound by scintillation counting, measuring the amount of radiolabeled first substrate reacted in the absence of the test compound by scintillation counting, and comparing the two measurements; and e. wherein when the amount of reacted first substrate is lower in the presence of a test compound than in the absence of the test compound, the test compound is identified as an inhibitor.