The interaction of myelin basic protein with cholesterol and the conformational changes occurring in the protein upon interaction with the lipid were investigated. The myelin basic protein (MBP) plays an important role in stabilizing the multilamellar structure of the myelin membrane. MBP interacts in a specific way with the lipids components of the membrane. The major lipid component is the cholesterol which comprises 40-44 mol% of the lipids. In order to understand the effect of the lipids in the protein conformation we have studied the interaction between MBP and cholesterol. The conformational changes induced in the protein upon interaction with different concentrations of cholesterol were characterized by transmission electron microscopy (TEM) and monolayer studies. Aqueous solution of MBP from bovine brain (obtained by the method of Cheifetz and Moscarello) exhibited a circular dichroism (CD) spectrum characteristic of random coil protein molecules. Upon addition of cholesterol, MBP-cholesterol complexes were observed by TEM. The monolayer compression experiments show plateaus in their surface pressure-area isotherms. The presence of these plateaus has previously been interpreted as ac-helix conformation. By seeding the MBP onto the aqueous support, we have determined the compression work for the protein on the surface. Experimental areas of the mixtures MBP-cholesterol are smaller than the area calculated by adding the areas of the pure components, indicating that there are attractive forces between both components. The calculated entropy of compression indicates that the highest organization is reached when lipid and protein are almost in the same proportion,