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ICEKAT: an interactive online tool for calculating initial rates from continuous enzyme kinetic traces
Michael D Olp1, Kelsey S Kalous1, Brian C Smith2
1Department of Biochemistry, Medical College of Wisconsin, 8701 Watertown Plank Rd., Milwaukee, 53226, USA.
BMC Bioinformatics
|May 16, 2020
Summary
A new tool, ICEKAT, simplifies initial rate analysis for continuous enzyme kinetic assays. This semi-automated software aids in high-throughput screening and kinetic parameter determination, improving efficiency and accuracy.
Area of Science:
- Enzymology
- Biochemistry
- Pharmacology
Background:
- Continuous enzyme kinetic assays are crucial for high-throughput screening, offering rapid data acquisition.
- Manual analysis of kinetic traces is time-consuming and prone to errors, hindering efficiency.
- Existing software often lacks simplicity for rapid initial rate determination in continuous assays.
Purpose of the Study:
- To develop a user-friendly tool for semi-automated analysis of initial rates from continuous enzyme kinetic data.
- To facilitate the evaluation of Michaelis-Menten and EC50/IC50 kinetic parameters.
- To streamline data analysis for high-throughput screening (HTS) assays.
Main Methods:
- Development of an Interactive Continuous Enzyme Kinetics Analysis Tool (ICEKAT).
- Browser-based interactive fitting tools for defining kinetic trace ranges.
- Semi-automated calculation of initial rates and kinetic parameters.
Main Results:
- ICEKAT enables semi-automated initial rate calculation from continuous enzyme kinetic traces.
- The tool simplifies fitting procedures compared to general-purpose software like Excel and Graphpad Prism.
- Successfully analyzed over 500 kinetic traces for SIRT1 activity in a test case.
Conclusions:
- ICEKAT facilitates simultaneous visualization of initial rate fits and kinetic model parameters.
- The software serves as a valuable tool for enzymologists and a teaching aid for understanding kinetic parameter calculations.
- ICEKAT is freely available online to the research community.
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