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  • Title: Detection of cholesterol-rich microdomains in the inner leaflet of the plasma membrane.
    Author: Hayashi M, Shimada Y, Inomata M, Ohno-Iwashita Y.
    Journal: Biochem Biophys Res Commun; 2006 Dec 22; 351(3):713-8. PubMed ID: 17083918.
    Abstract:
    The C-terminal domain (D4) of perfringolysin O binds selectively to cholesterol in cholesterol-rich microdomains. To address the issue of whether cholesterol-rich microdomains exist in the inner leaflet of the plasma membrane, we expressed D4 as a fusion protein with EGFP in MEF cells. More than half of the EGFP-D4 expressed in stable cell clones was bound to membranes in raft fractions. Depletion of membrane cholesterol with beta-cyclodextrin reduced the amount of EGFP-D4 localized in raft fractions, confirming EGFP-D4 binding to cholesterol-rich microdomains. Subfractionation of the raft fractions showed most of the EGFP-D4 bound to the plasma membrane rather than to intracellular membranes. Taken together, these results strongly suggest the existence of cholesterol-rich microdomains in the inner leaflet of the plasma membrane.
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