Isolation and Partial Characterization of an Amphiphilic 56-kDa Fatty Acid Binding Protein from Rat Renal Basolateral Membrane

Abstract
We have identified a 56-kDa fatty acid binding protein in rat renal basolateral membrane and purified it by extraction in nonionic detergent (Triton X-100), followed by gel filtration, DEAE-cellulose chromatography, and affinity chromatography. The purified protein was homogeneous on polyacrylamide gel electrophoresis in the presence of Triton X-100 or SDS. It showed amphiphilic properties on gel filtration, polyacrylamide gel electrophoresis, and oleate-Sepharose 4B chromatography. Its molecular mass was estimated to be 56 kDa by SDS-polyacrylamide gel electrophoresis. The protein showed optimal binding activity at pH 7.5 and 37°C. The apparent Kd for palmitic acid was 0.79 μM. It was immunologically clearly distinct from renal cytosolic fatty acid binding protein.