ICEKAT: an interactive online tool for calculating initial rates from continuous enzyme kinetic traces

摘要

Continuous enzyme kinetic assays allow rapid acquisition of large amounts of kinetic traces. Therefore, data analysis often becomes the bottleneck of high-throughput enzyme kinetic assays. In cases where IC /EC values or the Michaelis-Menten parameters (or ) and are of principle interest, reduction of kinetic traces to initial rates avoids error arising from assumptions involved in analyzing the entire kinetic trace [ ]. The two primary methods for determining initial rates from kinetic traces are estimation of the early linear portion of the curve and methods using integrated forms of kinetic equations [ – ]. Currently available programs such as FITSIM [ ], DYNAFIT [ ], ENZO [ ], PCAT [ ], and KinTek offer sophisticated routines for fitting kinetic traces. These programs are useful for selecting among complex enzymatic models and analyzing experiments carried out under conditions that may not satisfy the assumptions associated with Michaelis-Menten kinetics [ , ], for example measuring catalysis inside cells. However, the additional complexity offered by these programs is often not required when analyzing in vitro experiments, making them inefficient and unnecessarily complicated for many continuous enzyme kinetic applications.