Endocytosis of formaldehyde-treated serum albumin (f-albumin) in isolated liver sinusoidal endothelial cells was studied. Uptake occurs via the scavenger receptor and was found to be very sensitive to the ionophore monensin. Binding at 4 degrees C of f-albumin was reduced to 50% of control values by preincubation for 2 min with 2 microM monensin. Both uptake and degradation of f-albumin were more sensitive to monensin. No lag-phase in the inhibitory effect on uptake and degradation was detected. A concentration of 0.1 microM monensin reduced uptake of f-albumin by 50%. Degradation of internalized f-albumin was reduced by 50% in the presence of 0.2 microM monensin. Since uptake and degradation of f-albumin were very sensitive to monensin, the effect of introducing the drug during endocytosis of the ligand was tested. All processing of f-albumin stopped instantly upon addition of monensin; hence, there seems to be no step in the endocytic process beyond which monensin is ineffective. The data suggest that the scavenger receptor of liver endothelial cells is internalized and recycled very rapidly.