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

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Lipids

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 (K m, V m, and k o) were determined from a fitted Michaelis-Menten-type equation modified by a term for a saturable (linear) component of the measured total uptake. V m values were different for some of the FA studied, and K m 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 V m 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.

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Abbreviations

FABP:

fatty acid-binding protein

IMEM-Zo:

improved minimal essential medium-zinc optional

MEM:

minimal essential medium

UFA:

uncomplexed FA

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Authors and Affiliations

  1. INIBIOLP (Instituto de Investigaciones Bioquímicas de La Plata), CONICET-UNLP, Facultad de Ciencias Médicas, Universidad Nacional de La Plata, 60 y 120 (1900), La Plata, Argentina

    Carlos A. Marra

  2. Departamento de Bioquímica y Biología Molecular, Facultad de Farmacia, Universidad de Granada, Campus Cartuja, Granada, España

    María Dolores Girón & María Dolores Suáre

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  1. Carlos A. Marra
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  2. María Dolores Girón
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  3. María Dolores Suáre
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Correspondence to Carlos A. Marra.

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Marra, C.A., Girón, M.D. & Suáre, M.D. Evidence in favor of a facilitated transport system for FA uptake in cultured L6 cells. Lipids 37, 273–283 (2002). https://doi.org/10.1007/s11745-002-0891-5

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  • DOI: https://doi.org/10.1007/s11745-002-0891-5

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