Binding of 13-HODE and 15-HETE to phospholipid bilayers, albumin, and intracellular fatty acid binding proteins. implications for transmembrane and intracellular transport and for protection from lipid peroxidation

• Published in: The Journal of biological chemistry 2001-05, Vol.276 (19), p.15575-15580
• Main Authors: Ek-Von Mentzer, B A, Zhang, F, Hamilton, J A
• Format: Article
• Language: English
• Subjects: Animals; Binding Sites; Carrier Proteins - chemistry; Carrier Proteins - metabolism; Cattle; Fatty Acid-Binding Protein 7; Fatty Acid-Binding Proteins; Fatty Acids, Nonesterified - chemistry; Fatty Acids, Nonesterified - metabolism; Fatty Acids, Unsaturated - chemistry; Fatty Acids, Unsaturated - metabolism; Fluorescent Dyes; Hydroxyeicosatetraenoic Acids - chemistry; Hydroxyeicosatetraenoic Acids - metabolism; Kinetics; Linoleic Acids - chemistry; Linoleic Acids - metabolism; Lipid Bilayers - chemistry; Lipid Bilayers - metabolism; Lipid Peroxidation; Neoplasm Proteins; Nerve Tissue Proteins; Oxidation-Reduction; Phosphatidylcholines - chemistry; Phosphatidylcholines - metabolism; Protein Transport; Rats; Receptors, Retinoic Acid - chemistry; Receptors, Retinoic Acid - metabolism; Recombinant Proteins - chemistry; Recombinant Proteins - metabolism; Serum Albumin, Bovine - chemistry; Serum Albumin, Bovine - metabolism; Spectrometry, Fluorescence
• ISSN: 0021-9258
• DOI: 10.1074/jbc.M011623200

Transport and utilization of fatty acids (FA) in cells is a multistep process that includes adsorption to and movement across the plasma membrane and binding to intracellular fatty acid binding proteins (FABP) in the cytosol. We monitored the transbilayer movement of several polyunsaturated FA and oxidation products (13-hydroxy octadecadienoic acid (HODE) and 15-hydroxytetraenoic acid (HETE)) in unilamellar protein-free phospholipid vesicles containing a fluorescent pH probe. All FA diffused rapidly by the flip-flop mechanism across the model membrane, as revealed by pH changes inside the vesicle. This result suggests that FA oxidation products generated in the cell could cross the plasma or nuclear membrane spontaneously without a membrane transporter. To illuminate features of extra- and intracellular transport, the partitioning of unsaturated FA and oxidized FA between phospholipid vesicles and albumin or FABP was studied by the pyranin assay. These experiments showed that all polyunsaturated FA and oxidized FA (13-HODE and 15-HETE) desorbed rapidly from the phospholipid bilayer to bind to bovine serum albumin, which showed a slight preference for the unsaturated FA over the oxidized FA. FABP rapidly bound FA in the presence of phospholipid bilayers, with a preference of 13-HODE over the unsaturated FA and with a specificity depending on the type of FABP. Liver FABP was significantly more effective than intestinal FABP in binding 13-HODE in the presence of vesicles. The more effective binding of the FA metabolite, 13-HODE, than its precursor 18:2 by FABP may help protect cellular membranes from potential damage by monohydroxy fatty acids and may contribute a pathway for entry of 13-HODE into the nucleus.