Organization of fluorescent cholesterol analogs in lipid bilayers — Lessons from cyclodextrin extraction

Sigrid Milles; Thomas Meyer; Holger A. Scheidt; Roland Schwarzer; Lars  Thomas; Magdalena Marek; Lajos Szente; Robert Bittman; Andreas Herrmann; Thomas Günther-Pomorski; Daniel Huster; Peter Müller

doi:10.1016/j.bbamem.2013.04.002

Organization of fluorescent cholesterol analogs in lipid bilayers — Lessons from cyclodextrin extraction

Sigrid Milles, Thomas Meyer, Holger A. Scheidt, Roland Schwarzer, Lars Thomas, Magdalena Marek, Lajos Szente, Robert Bittman, Andreas Herrmann, Thomas Günther-Pomorski, Daniel Huster, Peter Müller

Section for Transport Biology

34 Citations (Scopus)

Abstract

To characterize the structure and dynamics of cholesterol in membranes, fluorescent analogs of the native molecule have widely been employed. The cholesterol content in membranes is in general manipulated by using water-soluble cyclodextrins. Since the interactions between cyclodextrins and fluorescent-labeled cholesterol have not been investigated in detail so far, we have compared the cyclodextrin-mediated membrane extraction of three different fluorescent cholesterol analogs (one bearing a NBD and two bearing BODIPY moieties). Extraction of these analogs was followed by measuring the Förster resonance energy transfer between a rhodamine moiety linked to phosphatidylethanolamine and the labeled cholesterol. The extraction kinetics revealed that the analogs are differently extracted from membranes. We examined the orientation of the analogs within the membrane and their influence on lipid condensation using NMR and EPR spectroscopies. Our data indicate that the extraction of fluorescent sterols from membranes is determined by several parameters, including their impact on lipid order, their hydrophobicity, their intermolecular interactions with surrounding lipids, their orientation within the bilayer, and their affinity with the exogenous acceptor.

Original language	English
Journal	B B A - Biomembranes
Volume	1828
Issue number	8
Pages (from-to)	1822-1828
Number of pages	7
ISSN	0005-2736
DOIs	https://doi.org/10.1016/j.bbamem.2013.04.002
Publication status	Published - 2013

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@article{e448928aed59419c83a626a936a88446,

title = "Organization of fluorescent cholesterol analogs in lipid bilayers — Lessons from cyclodextrin extraction",

abstract = "To characterize the structure and dynamics of cholesterol in membranes, fluorescent analogs of the native molecule have widely been employed. The cholesterol content in membranes is in general manipulated by using water-soluble cyclodextrins. Since the interactions between cyclodextrins and fluorescent-labeled cholesterol have not been investigated in detail so far, we have compared the cyclodextrin-mediated membrane extraction of three different fluorescent cholesterol analogs (one bearing a NBD and two bearing BODIPY moieties). Extraction of these analogs was followed by measuring the F{\"o}rster resonance energy transfer between a rhodamine moiety linked to phosphatidylethanolamine and the labeled cholesterol. The extraction kinetics revealed that the analogs are differently extracted from membranes. We examined the orientation of the analogs within the membrane and their influence on lipid condensation using NMR and EPR spectroscopies. Our data indicate that the extraction of fluorescent sterols from membranes is determined by several parameters, including their impact on lipid order, their hydrophobicity, their intermolecular interactions with surrounding lipids, their orientation within the bilayer, and their affinity with the exogenous acceptor.",

author = "Sigrid Milles and Thomas Meyer and Scheidt, {Holger A.} and Roland Schwarzer and Lars Thomas and Magdalena Marek and Lajos Szente and Robert Bittman and Andreas Herrmann and Thomas G{\"u}nther-Pomorski and Daniel Huster and Peter M{\"u}ller",

year = "2013",

doi = "10.1016/j.bbamem.2013.04.002",

language = "English",

volume = "1828",

pages = "1822--1828",

journal = "B B A - Biomembranes",

issn = "0005-2736",

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number = "8",

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TY - JOUR

T1 - Organization of fluorescent cholesterol analogs in lipid bilayers — Lessons from cyclodextrin extraction

AU - Milles, Sigrid

AU - Meyer, Thomas

AU - Scheidt, Holger A.

AU - Schwarzer, Roland

AU - Thomas, Lars

AU - Marek, Magdalena

AU - Szente, Lajos

AU - Bittman, Robert

AU - Herrmann, Andreas

AU - Günther-Pomorski, Thomas

AU - Huster, Daniel

AU - Müller, Peter

PY - 2013

Y1 - 2013

N2 - To characterize the structure and dynamics of cholesterol in membranes, fluorescent analogs of the native molecule have widely been employed. The cholesterol content in membranes is in general manipulated by using water-soluble cyclodextrins. Since the interactions between cyclodextrins and fluorescent-labeled cholesterol have not been investigated in detail so far, we have compared the cyclodextrin-mediated membrane extraction of three different fluorescent cholesterol analogs (one bearing a NBD and two bearing BODIPY moieties). Extraction of these analogs was followed by measuring the Förster resonance energy transfer between a rhodamine moiety linked to phosphatidylethanolamine and the labeled cholesterol. The extraction kinetics revealed that the analogs are differently extracted from membranes. We examined the orientation of the analogs within the membrane and their influence on lipid condensation using NMR and EPR spectroscopies. Our data indicate that the extraction of fluorescent sterols from membranes is determined by several parameters, including their impact on lipid order, their hydrophobicity, their intermolecular interactions with surrounding lipids, their orientation within the bilayer, and their affinity with the exogenous acceptor.

AB - To characterize the structure and dynamics of cholesterol in membranes, fluorescent analogs of the native molecule have widely been employed. The cholesterol content in membranes is in general manipulated by using water-soluble cyclodextrins. Since the interactions between cyclodextrins and fluorescent-labeled cholesterol have not been investigated in detail so far, we have compared the cyclodextrin-mediated membrane extraction of three different fluorescent cholesterol analogs (one bearing a NBD and two bearing BODIPY moieties). Extraction of these analogs was followed by measuring the Förster resonance energy transfer between a rhodamine moiety linked to phosphatidylethanolamine and the labeled cholesterol. The extraction kinetics revealed that the analogs are differently extracted from membranes. We examined the orientation of the analogs within the membrane and their influence on lipid condensation using NMR and EPR spectroscopies. Our data indicate that the extraction of fluorescent sterols from membranes is determined by several parameters, including their impact on lipid order, their hydrophobicity, their intermolecular interactions with surrounding lipids, their orientation within the bilayer, and their affinity with the exogenous acceptor.

U2 - 10.1016/j.bbamem.2013.04.002

DO - 10.1016/j.bbamem.2013.04.002

M3 - Journal article

C2 - 23583923

SN - 0005-2736

VL - 1828

SP - 1822

EP - 1828

JO - B B A - Biomembranes

JF - B B A - Biomembranes

IS - 8

ER -

Organization of fluorescent cholesterol analogs in lipid bilayers — Lessons from cyclodextrin extraction

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