Mechanism of free fatty acid transfer from rat heart fatty acid-binding protein to phospholipid membranes. Evidence for a collisional process.

Abstract

Fatty acid binding proteins (FABP) are a family of low molecular weight proteins found in many tissues that actively utilize free fatty acids (ffa). FABP would be expected to have a particularly important role in the heart, where over 80% of energy requirements are derived from oxidation of long chain fatty acids. The precise physiological function of heart FABP (H-FABP) has not been definitively identified, although it is thought to play a role in intracellular ffa transport. To examine the possible role of H-FABP in cardiac myocyte transfer of ffa, we examined the transfer of fluorescent anthroyloxy ffa (AOffa) from H-FABP to model phospholipid membranes, using a resonance energy transfer assay. In contrast to previous observations of ffa transfer from liver FABP and from membranes, transfer from H-FABP to membranes appears to occur by a different mechanism. AO-palmitate (16:0) transfer was 1.5-fold slower than AO-stearate (18:0) transfer, and mono-unsaturation did not affect the transfer rate. The AOffa transfer rate from H-FABP increased with increasing ionic strength and decreased slightly between pH 7 and 9. These results suggest that the rate of ffa transfer from H-FABP to membranes is independent of the ffa aqueous solubility. Thermodynamic analysis showed that the free energy of activation for the ffa transfer process arises primarily from an enthalpic component, with only a small entropic contribution, again suggesting the lack of an aqueous phase route of ffa delivery. Finally, the ffa transfer rate was found to be directly dependent on the concentration of acceptor membranes. These data therefore suggest that transfer of AOffa from H-FABP to membranes may occur via collisional interactions between the protein and membranes.