We have investigated the thermodynamic properties of the cholesterol transfer reaction from various types of liposomes to purified steroid-free cytochrome P450scc. From the results of temperature effect on the reaction, we obtained delta G+, delta H+, and delta S+ values for the reaction with various phosphatidylcholine vesicles containing different fatty acyl chains. The plots of delta H+ against delta S+, and delta G+ against delta H+ both reveal that an enthalpy-entropy compensation effect was seen in a series of phospholipid and micellar media of cholesterol. Our beta-value of 420 degrees K is unusually high among reported values for biochemical reactions.