The effect of random measurement errors on kinetic transport parameter estimation.

Abstract

Saturation kinetics experiments, in which uptake U of a substance across a transport barrier is measured as a function of initial concentration difference C, are used to describe transport of nutrients. Many such processes are characterized by low- and high-affinity systems in which kinetic parameters Vmax and Km differ by orders of magnitude. Transformations of equations to straight-line relationships between U and C are popular methods of parameter estimation. The aims of this study are (1) to show effects of random errors in U measurement on Vmax and Km estimation in a two-affinity process under several transformations: Lineweaver-Burk (1/U vs. 1/C), Hanes (C/U vs. C), Eadie-Hofstee (U/C vs. U), and Wolff (U vs. U/C), and (2) to indicate strategies for minimizing effects of errors. Two transport properties will illustrate: an ideal process of low- (Vmax = 100, Km = 10) and high-affinity (Vmax = 1, Km = .1) systems to which random error is added, and experimental uptake of 5-methyltetrahydrofolic acid by isolated hepatocytes.