Analysis of interactions between proteins and fatty acids or cholesterol using a fatty acid/cholesterol pull-down assay

Abstract Specific interactions between the polar head groups of membrane lipids and proteins have been described previously. In contrast, the specificity of the interaction between lipid acyl chains with proteins is less understood. By combining a fatty acid or cholesterol pull-down assay with Western blot analysis or mass spectrometry, we identified transmembrane and cytosolic proteins that bound preferentially to short or long acyl chains or to cholesterol. Thus, this approach allows identification of specific fatty acid–protein or cholesterol–protein interactions.