title: Direct determination of enzyme kinetic parameters from single reactions using a new progress curve analysis tool.
abstract: With computer-based data-fitting methods becoming a standard tool in biochemistry, progress curve analysis of enzyme kinetics is a feasible, yet seldom used tool. Here we present a versatile Matlab-based tool (PCAT) to analyze catalysis progress curves with three complementary model approaches. The first two models are based on the known closed-form solution for this problem: the first describes the required Lambert W function with an analytical approximation and the second provides a numerical solution of the Lambert W function. The third model is a direct simulation of the enzyme kinetics. Depending on the chosen model, the tools excel in speed, accuracy or initial value requirements. Using simulated and experimental data, we show the strengths and pitfalls of the different fitting models. Direct simulation proves to have the highest level of accuracy, but it also requires reasonable initial values to converge. Finally, we propose a standard procedure to obtain optimized enzyme kinetic parameters from single progress curves.
authors.affiliation: Department of Biomolecular Sciences, Weizmann Institute of Science, 234 Herzel St, Rehovot 76100, Israel.|Max Planck Institute for Dynamics and Self-Organization, Am Faßberg 17, D-37077 Goettingen, Germany.
authors.forename: Felix
authors.initials: F
authors.lastname: Bäuerle
authors.affiliation: Department of Biomolecular Sciences, Weizmann Institute of Science, 234 Herzel St, Rehovot 76100, Israel.
authors.forename: Agnes
authors.initials: A
authors.lastname: Zotter
authors.affiliation: Department of Biomolecular Sciences, Weizmann Institute of Science, 234 Herzel St, Rehovot 76100, Israel.
authors.forename: Gideon
authors.initials: G
authors.lastname: Schreiber
mesh_terms: D004798:Enzymes / Q000737:chemistry*; D007700:Kinetics; D008956:Models, Chemical; D012984:Software
chemical_list: D004798:Enzymes
keywords: Michaelis–Menten; enzyme kinetics; single reaction curve