Fluorescence assays for measuring fatty acid binding and transport through membranes.

The authors' laboratory has applied a series of different fluorescence assays for monitoring the binding and transport of fatty acids (FA) in model and biological membranes. The authors recently expanded their fluorescent assays for monitoring the adsorption of FA to membranes to a total of three probes that measure different aspects of FA binding: (1) an acrylodan-labeled FA-binding protein, which measures the partitioning of FA between membranes and the aqueous buffer; (2) the naturally occurring fluorescent cis-parinaric acid, which specifically measures the insertion of the FA acyl chain into the hydrophobic core of the phospholipid bilayer, and (3) a fluorescein-labeled phospholipid (N-fluorescein-5-thiocarbomoyl-1,2,dihexadecanoyl-sn-glycero-3-phosphoethanolamine), which specifically measures the arrival of the FA carboxyl at the outer leaflet of the membrane. None of these probes allow the transmembrane movement of FA to the inner leaflet to be measured.