Accurate label-free reaction kinetics determination using initial rate heat measurements.

ABSTRACT: Accurate label-free methods or assays to obtain the initial reaction rates have significant importance in fundamental studies of enzymes and in application-oriented high throughput screening of enzyme activity. Here we introduce a label-free approach for obtaining initial rates of enzyme activity from heat measurements, which we name initial rate calorimetry (IrCal). This approach is based on our new finding that the data recorded by isothermal titration calorimetry for the early stages of a reaction, which have been widely ignored, are correlated to the initial rates. Application of the IrCal approach to various enzymes led to accurate enzyme kinetics parameters as compared to spectroscopic methods and enabled enzyme kinetic studies with natural substrate, e.g. proteases with protein subs

SUBMITTER: Ebrahimi KH 

PROVIDER: S-EPMC4647221 | biostudies-literature | 2015 Nov

REPOSITORIES: biostudies-literature