Membrane orientation and lateral diffusion of BODIPY-cholesterol as a function of probe structure

Lukasz Michal Solanko, Daniel Wüstner, Frederik Wendelboe Lund, Jonathan R. Brewer, Henrik Midtiby, Robert Bittman, Christian Eggeling, Alf Honigmann, Vjekoslav Dekaris

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Cholesterol tagged with the BODIPY fluorophore via the central difluoroboron moiety of the dye (B-Chol) is a promising probe for studying intracellular cholesterol dynamics. We synthesized a new BODIPY-cholesterol probe (B-P-Chol) with the fluorophore attached via one of its pyrrole rings to carbon-24 of cholesterol (B-P-Chol). Using two-photon fluorescence polarimetry in giant unilamellar vesicles and in the plasma membrane (PM) of living intact and actin-disrupted cells, we show that the BODIPY-groups in B-Chol and B-P-Chol are oriented perpendicular and almost parallel to the bilayer normal, respectively. B-Chol is in all three membrane systems much stronger oriented than B-P-Chol. Interestingly, we found that the lateral diffusion in the PM was two times slower for B-Chol than for B-P-Chol, although we found no difference in lateral diffusion in model membranes. Stimulated emission depletion microscopy, performed for the first time, to our knowledge, with fluorescent sterols, revealed that the difference in lateral diffusion of the BODIPY-cholesterol probes was not caused by anomalous subdiffusion, because diffusion of both analogs in the PM was free but not hindered. Our combined measurements show that the position and orientation of the BODIPY moiety in cholesterol analogs have a severe influence on lateral diffusion specifically in the PM of living cells.
OriginalsprogEngelsk
TidsskriftBiophysical Journal
Vol/bind105
Udgave nummer9
Sider (fra-til)2082-2092
Antal sider10
ISSN0006-3495
DOI
StatusUdgivet - 5. nov. 2013

Fingeraftryk

Cholesterol
Membranes
Cell membranes
Fluorophores
4,4-difluoro-4-bora-3a,4a-diaza-s-indacene
Unilamellar Liposomes
Stimulated emission
Pyrroles
Polarimeters
Sterols
Actins
Microscopic examination
Coloring Agents
Carbon
Photons
Fluorescence
Cells

Citer dette

Solanko, Lukasz Michal ; Wüstner, Daniel ; Lund, Frederik Wendelboe ; Brewer, Jonathan R. ; Midtiby, Henrik ; Bittman, Robert ; Eggeling, Christian ; Honigmann, Alf ; Dekaris, Vjekoslav . / Membrane orientation and lateral diffusion of BODIPY-cholesterol as a function of probe structure. I: Biophysical Journal. 2013 ; Bind 105, Nr. 9. s. 2082-2092.
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abstract = "Cholesterol tagged with the BODIPY fluorophore via the central difluoroboron moiety of the dye (B-Chol) is a promising probe for studying intracellular cholesterol dynamics. We synthesized a new BODIPY-cholesterol probe (B-P-Chol) with the fluorophore attached via one of its pyrrole rings to carbon-24 of cholesterol (B-P-Chol). Using two-photon fluorescence polarimetry in giant unilamellar vesicles and in the plasma membrane (PM) of living intact and actin-disrupted cells, we show that the BODIPY-groups in B-Chol and B-P-Chol are oriented perpendicular and almost parallel to the bilayer normal, respectively. B-Chol is in all three membrane systems much stronger oriented than B-P-Chol. Interestingly, we found that the lateral diffusion in the PM was two times slower for B-Chol than for B-P-Chol, although we found no difference in lateral diffusion in model membranes. Stimulated emission depletion microscopy, performed for the first time, to our knowledge, with fluorescent sterols, revealed that the difference in lateral diffusion of the BODIPY-cholesterol probes was not caused by anomalous subdiffusion, because diffusion of both analogs in the PM was free but not hindered. Our combined measurements show that the position and orientation of the BODIPY moiety in cholesterol analogs have a severe influence on lateral diffusion specifically in the PM of living cells.",
author = "Solanko, {Lukasz Michal} and Daniel W{\"u}stner and Lund, {Frederik Wendelboe} and Brewer, {Jonathan R.} and Henrik Midtiby and Robert Bittman and Christian Eggeling and Alf Honigmann and Vjekoslav Dekaris",
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Membrane orientation and lateral diffusion of BODIPY-cholesterol as a function of probe structure. / Solanko, Lukasz Michal; Wüstner, Daniel; Lund, Frederik Wendelboe; Brewer, Jonathan R.; Midtiby, Henrik; Bittman, Robert; Eggeling, Christian; Honigmann, Alf; Dekaris, Vjekoslav .

I: Biophysical Journal, Bind 105, Nr. 9, 05.11.2013, s. 2082-2092.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Membrane orientation and lateral diffusion of BODIPY-cholesterol as a function of probe structure

AU - Solanko, Lukasz Michal

AU - Wüstner, Daniel

AU - Lund, Frederik Wendelboe

AU - Brewer, Jonathan R.

AU - Midtiby, Henrik

AU - Bittman, Robert

AU - Eggeling, Christian

AU - Honigmann, Alf

AU - Dekaris, Vjekoslav

PY - 2013/11/5

Y1 - 2013/11/5

N2 - Cholesterol tagged with the BODIPY fluorophore via the central difluoroboron moiety of the dye (B-Chol) is a promising probe for studying intracellular cholesterol dynamics. We synthesized a new BODIPY-cholesterol probe (B-P-Chol) with the fluorophore attached via one of its pyrrole rings to carbon-24 of cholesterol (B-P-Chol). Using two-photon fluorescence polarimetry in giant unilamellar vesicles and in the plasma membrane (PM) of living intact and actin-disrupted cells, we show that the BODIPY-groups in B-Chol and B-P-Chol are oriented perpendicular and almost parallel to the bilayer normal, respectively. B-Chol is in all three membrane systems much stronger oriented than B-P-Chol. Interestingly, we found that the lateral diffusion in the PM was two times slower for B-Chol than for B-P-Chol, although we found no difference in lateral diffusion in model membranes. Stimulated emission depletion microscopy, performed for the first time, to our knowledge, with fluorescent sterols, revealed that the difference in lateral diffusion of the BODIPY-cholesterol probes was not caused by anomalous subdiffusion, because diffusion of both analogs in the PM was free but not hindered. Our combined measurements show that the position and orientation of the BODIPY moiety in cholesterol analogs have a severe influence on lateral diffusion specifically in the PM of living cells.

AB - Cholesterol tagged with the BODIPY fluorophore via the central difluoroboron moiety of the dye (B-Chol) is a promising probe for studying intracellular cholesterol dynamics. We synthesized a new BODIPY-cholesterol probe (B-P-Chol) with the fluorophore attached via one of its pyrrole rings to carbon-24 of cholesterol (B-P-Chol). Using two-photon fluorescence polarimetry in giant unilamellar vesicles and in the plasma membrane (PM) of living intact and actin-disrupted cells, we show that the BODIPY-groups in B-Chol and B-P-Chol are oriented perpendicular and almost parallel to the bilayer normal, respectively. B-Chol is in all three membrane systems much stronger oriented than B-P-Chol. Interestingly, we found that the lateral diffusion in the PM was two times slower for B-Chol than for B-P-Chol, although we found no difference in lateral diffusion in model membranes. Stimulated emission depletion microscopy, performed for the first time, to our knowledge, with fluorescent sterols, revealed that the difference in lateral diffusion of the BODIPY-cholesterol probes was not caused by anomalous subdiffusion, because diffusion of both analogs in the PM was free but not hindered. Our combined measurements show that the position and orientation of the BODIPY moiety in cholesterol analogs have a severe influence on lateral diffusion specifically in the PM of living cells.

U2 - 10.1016/j.bpj.2013.09.031

DO - 10.1016/j.bpj.2013.09.031

M3 - Journal article

VL - 105

SP - 2082

EP - 2092

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

IS - 9

ER -