Steroid hormones such as progesterone, testosterone, and estradiol are derived from cholesterol, a major constituent of biomembranes. Although the hormones might be expected to associate with the bilayer in a fashion similar to that of cholesterol, their biological action in regulating transcription of target genes involves transbilayer transfer by free diffusion, which is not observed for cholesterol. We used a novel combination of a continuum-solvent model and the downhill simplex search method for the calculation of the free energy of interaction of these hormones with lipid membranes, and compared these values to that of cholesterol-membrane interaction. The hormones were represented in atomic detail and the membrane as a structureless hydrophobic slab embedded in implicit water. A deep free-energy minimum of ∼-15 kcal/mol was obtained for cholesterol at its most favorable location in the membrane, whereas the most favorable locations for the hormones were associated with shallower minima of -5.0 kcal/mol or higher. The free-energy difference, which is predominantly due to the substitution of cholesterol's hydrophobic tail with polar groups, explains the different manner in which cholesterol and the hormones interact with the membrane. Further calculations were conducted to estimate the rate of transfer of the hormones from the aqueous phase into hexane, and from hexane back into the aqueous phase. The calculated rates agreed reasonably well with measurements in closely related systems. Based on these calculations, we suggest putative pathways for the free diffusion of the hormones across biomembranes. Overall, the calculations imply that the hormones may rapidly cross biomembrane barriers. Implications for gastrointestinal absorption and transfer across the blood-brain barrier and for therapeutic uses are discussed.