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Title: Detection of cholesterol-rich microdomains in the inner leaflet of the plasma membrane

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.

Authors:
 [1];  [1];  [1];  [1]
  1. Cellular Signaling Group, Research Team for Functional Genomics, Tokyo Metropolitan Institute of Gerontology, 35-2 Sakae-cho, Itabashi-ku, Tokyo 173-0015 (Japan)
Publication Date:
OSTI Identifier:
20857929
Resource Type:
Journal Article
Journal Name:
Biochemical and Biophysical Research Communications
Additional Journal Information:
Journal Volume: 351; Journal Issue: 3; Other Information: DOI: 10.1016/j.bbrc.2006.10.088; PII: S0006-291X(06)02357-6; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0006-291X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; CELL MEMBRANES; CHOLESTEROL; FIBROBLASTS; LIPIDS; MICE; PROTEINS

Citation Formats

Hayashi, Masami, Shimada, Yukiko, Inomata, Mitsushi, and Ohno-Iwashita, Yoshiko. Detection of cholesterol-rich microdomains in the inner leaflet of the plasma membrane. United States: N. p., 2006. Web. doi:10.1016/j.bbrc.2006.10.088.
Hayashi, Masami, Shimada, Yukiko, Inomata, Mitsushi, & Ohno-Iwashita, Yoshiko. Detection of cholesterol-rich microdomains in the inner leaflet of the plasma membrane. United States. doi:10.1016/j.bbrc.2006.10.088.
Hayashi, Masami, Shimada, Yukiko, Inomata, Mitsushi, and Ohno-Iwashita, Yoshiko. Fri . "Detection of cholesterol-rich microdomains in the inner leaflet of the plasma membrane". United States. doi:10.1016/j.bbrc.2006.10.088.
@article{osti_20857929,
title = {Detection of cholesterol-rich microdomains in the inner leaflet of the plasma membrane},
author = {Hayashi, Masami and Shimada, Yukiko and Inomata, Mitsushi and Ohno-Iwashita, Yoshiko},
abstractNote = {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.},
doi = {10.1016/j.bbrc.2006.10.088},
journal = {Biochemical and Biophysical Research Communications},
issn = {0006-291X},
number = 3,
volume = 351,
place = {United States},
year = {2006},
month = {12}
}