Exposure of phosphatidylinositol transfer proteins to sphingomyelin-cholesterol membranes suggests transient but productive interactions with raft-like, liquid-ordered domains.

Both isoforms of rat phosphatidylinositol transfer protein (PITP) mediate the intermembrane transfer of sphingomyelin (CerPCho). In the plasma membrane, CerPCho often segregates with cholesterol into microdomains such as lipid rafts and caveolae. To test the hypothesis that PITP exhibits a preference for CerPCho- and cholesterol-rich membranes, we prepared unilamellar vesicles containing variable amounts of these two lipids. We also used CerPCho species with different acyl composition and treated vesicles with agents known to sequester and remove cholesterol. We observed that the beta isoform of rat PITP was more sensitive to membrane cholesterol than was the alpha isoform, as shown by increases in specific activities of lipid transfer of 2-6-fold. A relatively high membrane content of cholesterol (mole fraction > 0.4) was required to elicit such enhancements. Treatment of cholesterol-rich membranes with a series of beta cyclodextrins demonstrated that, upon depletion of cholesterol from participating membranes, the PITPbeta activity changes were fully reversible. We finally noted that the mechanism by which cholesterol enhances the activity of PITPbeta appeared to involve a decreased affinity of the protein for the membrane surface, in a manner that was independent of vesicle size and membrane microviscosity. We conclude that PITPbeta interacts transiently but productively with the liquid-ordered phase formed by CerPCho and cholesterol and discuss the possibility of PITP interactions in vivo with sphingolipid- and cholesterol-rich membrane microdomains.