Addition of increasing amounts of benzyl alcohol progressively reduced the steady-state anisotropies of diphenylhexatriene and trimethylammoniumdiphenylhexatriene in brush-border membranes from rat kidney. The decrease in order of membrane lipids, equivalent for 50 mM benzyl alcohol to that produced by a rise in temperature of approx. 6 degrees C, had no effect on the activities of alkaline phosphatase or gamma-glutamyltranspeptidase. On the other hand, benzyl alcohol markedly inhibited the D-glucose uptakes measured in the presence of a 100 mM sodium gradient. For concentrations less than 30 mM, benzyl alcohol reduced the Jmax without significant effects on Km, 22Na+ uptake or the vesicular volume of brush-border preparations. Comparable results were obtained substituting octanol for benzyl alcohol. Our data strongly suggest that, at constant temperature, the D-glucose carrier present in renal brush-border membranes is extremely sensitive to variations in membrane physical state.