Directly imaging emergence of phase separation in peroxidized lipid membranes

Paez-Perez, Miguel; Vyšniauskas, Aurimas; López‐Duarte, Ismael; Lafarge, Eulalie; Castro, Raquel López-Ríos De; Marques, Carlos M.; Schröder, A.; Muller, Pierre; Lorenz, Christian D.; Brooks, Nicholas J.; Kuimova, Marina K.

doi:10.1038/s42004-022-00809-x

Cited by 25 publications

(27 citation statements)

References 87 publications

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“…Lipid peroxidation generates products such as truncated lipids that perturb lipid packing in synthetic model membranes. ,,− ,, To test whether similar effects are relevant in biological membranes, we turned to environmentally sensitive fluorescent probes that sense lipid packing. Specifically, we monitored the lifetime of the fluorescent reporter Di4 (Di-4-ANEPPDHQ), , which has been widely shown to be dependent on membrane lipid packing, with lower Di4 lifetimes indicative of looser packing and more fluid membranes, and vice versa …”

Section: Resultsmentioning

confidence: 99%

Lipid Peroxidation Drives Liquid–Liquid Phase Separation and Disrupts Raft Protein Partitioning in Biological Membranes

Balakrishnan,

Kenworthy

2024

J. Am. Chem. Soc.

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The peroxidation of membrane lipids by free radicals contributes to aging, numerous diseases, and ferroptosis, an iron-dependent form of cell death. Peroxidation changes the structure and physicochemical properties of lipids, leading to bilayer thinning, altered fluidity, and increased permeability of membranes in model systems. Whether and how lipid peroxidation impacts the lateral organization of proteins and lipids in biological membranes, however, remains poorly understood. Here, we employ cell-derived giant plasma membrane vesicles (GPMVs) as a model to investigate the impact of lipid peroxidation on ordered membrane domains, often termed membrane rafts. We show that lipid peroxidation induced by the Fenton reaction dramatically enhances the phase separation propensity of GPMVs into coexisting liquid-ordered (Lo) and liquid-disordered (Ld) domains and increases the relative abundance of the disordered phase. Peroxidation also leads to preferential accumulation of peroxidized lipids and 4-hydroxynonenal (4-HNE) adducts in the disordered phase, decreased lipid packing in both Lo and Ld domains, and translocation of multiple classes of raft proteins out of ordered domains. These findings indicate that the peroxidation of plasma membrane lipids disturbs many aspects of membrane rafts, including their stability, abundance, packing, and protein and lipid composition. We propose that these disruptions contribute to the pathological consequences of lipid peroxidation during aging and disease and thus serve as potential targets for therapeutic intervention.

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Section: Resultsmentioning

confidence: 99%

Lipid Peroxidation Drives Liquid–Liquid Phase Separation and Disrupts Raft Protein Partitioning in Biological Membranes

Balakrishnan,

Kenworthy

2024

J. Am. Chem. Soc.

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“…Lipid peroxidation generates products such as truncated lipids that perturb lipid packing in synthetic model membranes [10, 11, 14–16, 19, 20]. To test whether similar effects are relevant in biological membranes, we turned to environment-sensitive fluorescent probes that sense lipid packing.…”

Section: Resultsmentioning

confidence: 99%

“…Their hydroperoxidized acyl chains becomes more hydrophilic and tend to associate with the lipid-water interface [9][10][11]. Their presence in membranes ultimately increases membrane permeability and impacts other membranes properties such as packing, fluidity, and viscosity [12][13][14][15][16][17][18][19][20]. Further oxidation generates secondary products such as reactive aldehydes, ketones, alcohols and ethers.…”

Section: Introductionmentioning

confidence: 99%

Lipid peroxidation drives liquid-liquid phase separation and disrupts raft protein partitioning in biological membranes

Balakrishnan,

Kenworthy

2023

Preprint

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The peroxidation of membrane lipids by free radicals contributes to aging, numerous diseases, and ferroptosis, an iron-dependent form of cell death. Peroxidation changes the structure, conformation and physicochemical properties of lipids, leading to major membrane alterations including bilayer thinning, altered fluidity, and increased permeability. Whether and how lipid peroxidation impacts the lateral organization of proteins and lipids in biological membranes, however, remains poorly understood. Here, we employ cell-derived giant plasma membrane vesicles (GPMVs) as a model to investigate the impact of lipid peroxidation on ordered membrane domains, often termed membrane rafts. We show that lipid peroxidation induced by the Fenton reaction dramatically enhances phase separation propensity of GPMVs into co-existing liquid ordered (raft) and liquid disordered (non-raft) domains and increases the relative abundance of the disordered, non-raft phase. Peroxidation also leads to preferential accumulation of peroxidized lipids and 4-hydroxynonenal (4-HNE) adducts in the disordered phase, decreased lipid packing in both raft and non-raft domains, and translocation of multiple classes of proteins out of rafts. These findings indicate that peroxidation of plasma membrane lipids disturbs many aspects of membrane rafts, including their stability, abundance, packing, and protein and lipid composition. We propose that these disruptions contribute to the pathological consequences of lipid peroxidation during aging and disease, and thus serve as potential targets for therapeutic intervention.

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“…where 𝑧 and 𝛼 are calibration constants that are experimentally determined by measuring the MR lifetime in solutions of known viscosity. Importantly, the fluorescence lifetime is independent of the local probe concentration and instrument setup, and this allows the direct quantitative mapping of microviscosity in heterogeneous model 19,[21][22][23] and cellular [24][25][26] membranes under different stress conditions. 11,22,23,27 Therefore, we anticipate the fluorescence lifetime of membrane-embedded BODIPY-based MRs could be used to directly infer changes in the membrane's structure.…”

Section: 𝜏 = 𝑧𝜂 𝛼mentioning

confidence: 99%

Viscosity-sensitive membrane dyes as tools to estimate the crystalline structure of lipid bilayers

Paez-Perez

Dent

Brooks

et al. 2023

Preprint

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Lipid membranes are crucial for cellular integrity and regulation, and tight control of their structural and mechanical properties is vital to ensure that they function properly. Fluorescent probes sensitive to the mem-brane’s microenvironment are useful for investigating lipid membrane properties, however, there is currently a lack of quantitative correlation between the exact parameters of lipid organization and a readout from these dyes. Here, we investigate this relationship for ‘molecular rotors’, or microviscosity sensors, by simultaneously measuring their fluorescence lifetime to determine the membrane viscosity, while using the X-Ray diffraction the determine the membrane’s structural properties. Our results reveal a phase-dependent correlation be-tween the membrane’s structural parameters and mechanical properties measured by a BODIPY-based mo-lecular rotor, giving excellent predictive power for the structural descriptors of the lipid bilayer. We also demonstrate that differences in membrane thickness between different lipid phases is not a prerequisite for formation of lipid microdomains and that this requirement can be disrupted by the presence of line-active molecules. Our results underpin the use of membrane-sensitive dyes as reporters of the structure of lipid membranes.

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Directly imaging emergence of phase separation in peroxidized lipid membranes

Cited by 25 publications

References 87 publications

Lipid Peroxidation Drives Liquid–Liquid Phase Separation and Disrupts Raft Protein Partitioning in Biological Membranes

Lipid Peroxidation Drives Liquid–Liquid Phase Separation and Disrupts Raft Protein Partitioning in Biological Membranes

Lipid peroxidation drives liquid-liquid phase separation and disrupts raft protein partitioning in biological membranes

Viscosity-sensitive membrane dyes as tools to estimate the crystalline structure of lipid bilayers

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