Evidence in favor of a facilitated transport system for FA uptake in cultured L6 cells

Abstract

In this manuscript we report a study of the transport of FA in L6 muscle cells. Cultured L6 cells took up labeled FA (C10 to C20) as a linear function of time up to 15 min. Thereafter, the rate of uptake gradually declined although it persisted for at least 12 h after the addition of the substrate. Kinetic parameters (Km, Vm, and ko) were determined from a fitted Michaelis-Menten-type equation modified by a term for a saturable (linear) component of the measured total uptake. Vm values were different for some of the FA studied, and Km data showed significant differences between saturated and unsaturated FA. The maximal rate of uptake was observed at pH 7.40 for decanoate, palmitate, and eicosatrienoate. Uptake was significantly influenced when the pH of the incubation medium was changed. Experiments designed to study the influence of FA/albumin molar ratio indicated that Vm was dependent on the total (bound and free) concentration of the FA. A concentrative uptake was demonstrated in short-term experiments with an apparent plateau of 20 and 40 μM for palmitate and eicosatrienoate, respectively. A competitive inhibition was also observed between palmitate as substrate and the other FA. From our results we can postulate that the uptake of FA in L6 cells is the sum of passive diffusion plus a saturable component and that the rate of uptake is dependent on one (or more) protein structures, although their precise characteristics and functions remain to be elucidated.