Distribution and Transport of Cholesterol-rich Membrane Domains Monitored by a Membrane-impermeant Fluorescent Polyethylene Glycol-derivatized Cholesterol

Cholesterol-rich membrane domains function in various membrane events as diverse as signal transduction and membrane traffic. We studied the interaction of a fluorescein ester of polyethylene glycol-derivatized cholesterol (fPEG-Chol) with cholesterol-rich membranes both in cells and in model membra...

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Bibliographic Details
Published in:The Journal of biological chemistry 2004-05, Vol.279 (22), p.23790-23796
Main Authors: Sato, Satoshi B., Ishii, Kumiko, Makino, Asami, Iwabuchi, Kazuhisa, Yamaji-Hasegawa, Akiko, Senoh, Yukiko, Nagaoka, Isao, Sakuraba, Hitoshi, Kobayashi, Toshihide
Format: Article
Language:English
Subjects:
Biological Transport
Cells, Cultured
Cholesterol - analogs & derivatives
Cholesterol - metabolism
Fluorescent Dyes
Humans
Membrane Microdomains - chemistry
Membrane Microdomains - metabolism
Polyethylene Glycols
Sensitivity and Specificity
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Summary:Cholesterol-rich membrane domains function in various membrane events as diverse as signal transduction and membrane traffic. We studied the interaction of a fluorescein ester of polyethylene glycol-derivatized cholesterol (fPEG-Chol) with cholesterol-rich membranes both in cells and in model membranes. Unlike filipin and other cholesterol probes, this molecule could be applied as an aqueous dispersion to various samples. When added to live cells, fPEG-Chol distributed exclusively in the outer plasma membrane leaflet and was enriched in microdomains that dynamically clustered by the activation of receptor signaling. The surface-bound fPEG-Chol was slowly internalized via clathrin-independent pathway into endosomes together with lipid raft markers. Noteworthy, fPEG-Chol could be microinjected in the living cells in which we found Golgi apparatus as the sole major organelle to be labeled. PEG-Chol, thus, provides a novel, sensitive probe for unraveling the dynamics of cholesterol-rich microdomains in living cells.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M313568200