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Journal of Lipid Research, Vol. 42, 509-520, April 2001
Copyright © 2001 by Lipid Research, Inc.

Original Article

Oleic acid uptake and binding by rat adipocytes define dual pathways for cellular fatty acid uptake

Correspondence to: P. D. Berk, at the Division of Liver Disease (Box 1633), Mount Sinai School of Medicine, 1 Gustave L. Levy Place, New York, NY 10029., paul.berk{at}mssm.edu (E-mail)

Oleic acid (OA) uptake by rat adipocytes and the proportions of intracellular unesterified [³H]OA and its ³H-labeled esters were determined over 300 s. Uptake was linear for 20;–30 s, with rapid esterification indicating entry into normal metabolic pathways. Initial rates of OA uptake and its binding to plasma membranes were studied over a spectrum of oleic acid:bovine serum albumin (BSA) ratios, and expressed as functions of unbound OA concentrations calculated with both the 1971 OA:BSA association constants of Spector, Fletcher, and Ashbrook and more recent constants (e.g., the 1993 constants of Richieri, Anel, and Kleinfeld), which generate concentrations 10- to 100-fold lower. In either case, uptake was the sum of saturable and linear processes, with 90% occurring via the saturable pathway when the OA:BSA molar ratio was within the physiologic range (0.5;–3.0). Within this range, rate constants for saturable transmembrane influx (k_s), calculated from both sets of constants, were similar (2.9 s^-1) and were 10- to 30-fold faster than those for nonsaturable uptake (k_ns = 0.26;–0.10 s^-1, t_1/2 = 2.7;–6.6 s, based on the constants of Spector et al. and Richieri et al., respectively). The rate of oleic acid flip-flop into rat adipocytes (k_ff = 0.16 ± 0.02 s^-1, t_1/2 = 4.3 ± 0.5 s), computed from published data, was similar to k_ns.

Thus, OA uptake occurs by both a saturable mechanism and passive flip-flop. This conclusion is independent of the OA:BSA association constants used to analyze the experimental measurements. — Stump, D. D., X. Fan, and P. D. Berk. Oleic acid uptake and binding by rat adipocytes define dual pathways for cellular fatty acid uptake. J. Lipid Res. 2001. 42: 509;–520.

Supplementary key words: diffusion, facilitated transport, flip-flop, kinetics

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