Kinetics Workshop
An intensive, hands-on workshop covering the theory and practice of enzyme kinetics.
Program Content
Mechanism-based data fitting
The course will focus on the use of computer simulation based on numerical integration of rate equations obtained from a model entered by the user. The primary goal of the workshop will be to help you to become proficient in modeling and fitting data using KinTek Explorer software so that you can tackle any problem.
Steady state kinetics
The information content of steady state kinetic measurements will be described in discussing the meaning of the kinetic constants, what they tell about a reaction mechanism and what they do not reveal.
Introduction to transient kinetic methods
The need for transient kinetic methods will be described to provide mechanistic information that can be interpreted directly to define reactions occurring at the active sites of enzymes or nucleic acids.
Fundamental principles of reaction kinetics
The basic principles of reaction rate measurement will be described. The simple math behind exponential reaction kinetics will be presented to reveal the information content of kinetic data.
Data fitting principles and practice
The use of computer simulation and nonlinear regression in data fitting will be described. We will show how mechanism-based data fitting overcomes the many limitations of equation-based fitting.
Kinetics of ligand binding
We will begin a discussion of reaction kinetics with the binding of a ligand to a protein or nucleic acid to emphasize the importance of analysis of the concentration dependence of the rate.
Kinetics of multi-step reactions
The kinetics of two-step reactions will be described under different scenarios. The principles that govern the design of experiments and the modeling of data to distinguish alternatives will be discussed.
Global fitting methods
We will show how global data fitting based on computer simulation provides the most rigorous method to extract mechanistic information. This analysis reveals a complete pathway in ways that cannot be obtained by piecemeal equation-based data fitting.
Testimonials
"Thank you for organizing this intensive workshop with such passion and desire to share the knowledge you’ve acquired over the years. I’ll go back to the lab with a better and clearer understanding of how to treat my data, which is just a wonderful and valuable gift."
- Erika Lopez-Alfonzo, University of California Berkeley
"This workshop has been transformative in how I view experimental planning and analysis. It has been a great pleasure to attend."
- Michael Cory, University of Pennsylvania
Agenda
Tuesday, 9 Jan 2024 - Friday, 12 Jan 2024
- Day 1: Intro to Enzyme Kinetics, Computer Simulation I
- Day 2: Steady State Kinetics, Multi-step Reactions
- Day 3: Problem Solving, Stopped-Flow, Data Fitting
- Day 4: pH & Isotope Effects, Global Fitting, Single Molecule Kinetics