Cholesterol Depletion Induces Solid-like Regions in the Plasma Membrane

ABSTRACT Glycosylphosphatidylinositol-linked and transmembrane major histocompatibility complex (MHC) class II I-Ek proteins, as well as N-(6-tetramethylrhodaminethiocarbamoyl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine (Tritc-DHPE), are used as probes to determine the effect of cholesterol concentration on the organization of the plasma membrane at temperatures in the range 22C–42C. Cholesterol depletion caused a decrease in the diffusion coefficients for the MHC II proteins and also for a slow fraction of the Tritc-DHPE population. At 37C, reduction of the total cell cholesterol concentration results in a smaller suppression of the translational diffusion for I-Ek proteins (twofold) than was observed in earlier work at 22C (five sevenfold) Vrljic, M., S. Y. Nishimura, W. E. Moerner, and H. M. McConnell. 2005. Biophys. J. 88:334–347. At 37C, the diffusion of both I-Ek proteins is Brownian (0.9, a-parameter, 1.1). More than 99 % of the protein population diffuses homogeneously when imaged at 65 frames per s. As the temperature is raised from 22C to 42C, a change in activation energy is seen at;35C in the Arrhenius plots. Cytoskeletal effects appear to be minimal. These results are consistent with a previously described model of solid-like domain formation in the plasma membrane