Measurement of enzyme kinetics and inhibitor constants using enthalpy arrays
Enthalpy arrays enable label-free, solution-based calorimetric detection of molecular interactions in a 96-detector array format. The combination of the small size of the detectors and ability to perform measurements in parallel results in a significant reduction of sample volume and measurement time compared with conventional calorimetry. The current capabilities of the technology for studying enzyme-catalyzed reactions are demonstrated by determining the kinetic parameters for reactions with three model enzymes. In addition, the technology has been used with two classes of enzymes to determine accurate inhibitor constants for competitive inhibitors from measurements at a single inhibitor concentration. We also demonstrate noncompetitive inhibition to the second substrate with a two-substrate enzyme.
Recht, M. I.; Torres, F.; De Bruyker, D.; Bell, A. G.; Klumpp, M.; Bruce, R. H. Measurement of enzyme kinetics and inhibitor constants using enthalpy arrays. Analytical Biochemistry. 2009 May 15; 388 (2): 204-212.