Oxidized Phospholipids Inhibit the Formation of Cholesterol-Dependent Plasma Membrane Nanoplatforms

Mario Brameshuber, Eva Sevcsik, Benedikt K. Rossboth, Christina Manner, Hans Peter Deigner, Begüm Peksel, Mária Péter, Z. Török, Albin Hermetter, Gerhard J. Schütz

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Abstract

We previously developed a single-molecule microscopy method termed TOCCSL (thinning out clusters while conserving stoichiometry of labeling), which allows for direct imaging of stable nanoscopic platforms with raft-like properties diffusing in the plasma membrane. As a consensus raft marker, we chose monomeric GFP linked via a glycosylphosphatidylinositol (GPI) anchor to the cell membrane (mGFP-GPI). With this probe, we previously observed cholesterol-dependent homo-association to nanoplatforms diffusing in the plasma membrane of live CHO cells. Here, we report the release of this homo-association upon addition of 1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphocholine (POVPC) or 1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphocholine, two oxidized phospholipids (oxPLs) that are typically present in oxidatively modified low-density lipoprotein. We found a dose-response relationship for mGFP-GPI nanoplatform disintegration upon addition of POVPC, correlating with the signal of the apoptosis marker Annexin V-Cy3. Similar concentrations of lysolipid showed no effect, indicating that the observed phenomena were not linked to properties of the lipid bilayer itself. Inhibition of acid sphingomyelinase by NB-19 before addition of POVPC completely abolished nanoplatform disintegration by oxPLs. In conclusion, we were able to determine how oxidized lipid species disrupt mGFP-GPI nanoplatforms in the plasma membrane. Our results favor an indirect mechanism involving acid sphingomyelinase activity rather than a direct interaction of oxPLs with nanoplatform constituents.

Original languageEnglish
Pages (from-to)205-213
Number of pages9
JournalBiophysical Journal
Volume110
Issue number1
DOIs
Publication statusPublished - Jan 5 2016

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Glycosylphosphatidylinositols
Phospholipids
Cholesterol
Cell Membrane
Sphingomyelin Phosphodiesterase
Phosphorylcholine
Acids
CHO Cells
Annexin A5
Lipid Bilayers
LDL Lipoproteins
Apoptosis
Lipids

ASJC Scopus subject areas

  • Biophysics

Cite this

Brameshuber, M., Sevcsik, E., Rossboth, B. K., Manner, C., Deigner, H. P., Peksel, B., ... Schütz, G. J. (2016). Oxidized Phospholipids Inhibit the Formation of Cholesterol-Dependent Plasma Membrane Nanoplatforms. Biophysical Journal, 110(1), 205-213. https://doi.org/10.1016/j.bpj.2015.11.018

Oxidized Phospholipids Inhibit the Formation of Cholesterol-Dependent Plasma Membrane Nanoplatforms. / Brameshuber, Mario; Sevcsik, Eva; Rossboth, Benedikt K.; Manner, Christina; Deigner, Hans Peter; Peksel, Begüm; Péter, Mária; Török, Z.; Hermetter, Albin; Schütz, Gerhard J.

In: Biophysical Journal, Vol. 110, No. 1, 05.01.2016, p. 205-213.

Research output: Contribution to journalArticle

Brameshuber, M, Sevcsik, E, Rossboth, BK, Manner, C, Deigner, HP, Peksel, B, Péter, M, Török, Z, Hermetter, A & Schütz, GJ 2016, 'Oxidized Phospholipids Inhibit the Formation of Cholesterol-Dependent Plasma Membrane Nanoplatforms', Biophysical Journal, vol. 110, no. 1, pp. 205-213. https://doi.org/10.1016/j.bpj.2015.11.018
Brameshuber M, Sevcsik E, Rossboth BK, Manner C, Deigner HP, Peksel B et al. Oxidized Phospholipids Inhibit the Formation of Cholesterol-Dependent Plasma Membrane Nanoplatforms. Biophysical Journal. 2016 Jan 5;110(1):205-213. https://doi.org/10.1016/j.bpj.2015.11.018
Brameshuber, Mario ; Sevcsik, Eva ; Rossboth, Benedikt K. ; Manner, Christina ; Deigner, Hans Peter ; Peksel, Begüm ; Péter, Mária ; Török, Z. ; Hermetter, Albin ; Schütz, Gerhard J. / Oxidized Phospholipids Inhibit the Formation of Cholesterol-Dependent Plasma Membrane Nanoplatforms. In: Biophysical Journal. 2016 ; Vol. 110, No. 1. pp. 205-213.
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