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. Compared with conventional calorimetry, enthalpy arrays achieve a significant reduction of sample volume and measurement time through the combination of the small size of the detectors and ability to perform measurements in parallel. 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.
Recht
Michael I
MI
Palo Alto Research Center, Palo Alto, CA 94304, USA. mrecht@parc.com
Torres
Frank E
FE
De Bruyker
Dirk
D
Bell
Alan G
AG
Klumpp
Martin
M
Bruce
Richard H
RH
D002151:Calorimetry / Q000379:methods*; D004798:Enzymes / Q000378:metabolism*; D007700:Kinetics; D036103:Nanotechnology / Q000379:methods*
D004798:Enzymes