The surface properties of small intestine brush-border membranes (BBMs) were examined by frontal affinity chromatography using three types of unsolubilized ligands: phlorizin polymer, immobilized lectins and linolenic acid bound to agarose gel. BBM vesicles were purified from piglets fed a corn oil diet (control diet) or a hydrogenated coconut oil diet. The second diet was representative of a deficient supply of essential polyunsaturated fatty acid (EPUFA). It induced a marked decrease in 18:2n-6 content in membrane choline phosphoglycerides and ethanolamine phosphoglycerides, whereas 20:3n-9 appeared in each class of phospholipids. Control and EPUFA-deprived BBM vesicles bound to phlorizin polymer, linolenic acid-agarose and wheat germ agglutinin (WGA) gel. In contrast, concanavalin A gel and Lens culinaris A gel exhibited a low binding capacity towards the two types of vesicles. EPUFA deficiency induced a slight decrease in binding on phlorizin polymer and a marked increase in binding on WGA gel, whereas the two types of vesicles similarly bound to linolenic acid-agarose. Desorption of phlorizin polymer-bound membranes was performed using several detergents with special regard to sodium deoxycholate (NaDOC) micelles. Sucrase activity recovery showed that the efficiency of NaDOC desorption was diminished in the case of EPUFA-deprived vesicles. EPUFA-deprived membrane domains involved in the binding would be less sensitive to the detergent attack. This assumption agrees with the putative decrease in membrane fluidity induced by the deficient diet. The possibility that fatty acid compositional changes induced by dietary lipids are extensive enough to alter some chromatographic properties of BBM vesicles is discussed.