Dithiothreitol Raises Transition Temperatures in Giant Plasma Membrane Vesicles

Gerstle, Ruth; Desai, Rohan; Veatch, Sarah L.

doi:10.1016/j.bpj.2016.11.2808

Cited by 4 publications

(3 citation statements)

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“…For example, GPMVs prepared with dithiothreitol (DTT) and paraformaldehyde (PFA) exhibit a demixing temperature about 10–20 °C higher than that of GPMVs derived using N -ethylmaleimide (NEM) 26 . DTT has been shown to affect lipid–lipid and lipid–protein interactions and to integrate directly into lipid membranes 32,33 . To figure out potential differences in mechanical properties, the bending rigidity of GPMVs formed using these two most widely employed methods was assessed.…”

Section: Resultsmentioning

confidence: 99%

Mechanical properties of plasma membrane vesicles correlate with lipid order, viscosity and cell density

et al. 2019

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Regulation of plasma membrane curvature and composition governs essential cellular processes. The material property of bending rigidity describes the energetic cost of membrane deformations and depends on the plasma membrane molecular composition. Because of compositional fluctuations and active processes, it is challenging to measure it in intact cells. Here, we study the plasma membrane using giant plasma membrane vesicles (GPMVs), which largely preserve the plasma membrane lipidome and proteome. We show that the bending rigidity of plasma membranes under varied conditions is correlated to readout from environment-sensitive dyes, which are indicative of membrane order and microviscosity. This correlation holds across different cell lines, upon cholesterol depletion or enrichment of the plasma membrane, and variations in cell density. Thus, polarity- and viscosity-sensitive probes represent a promising indicator of membrane mechanical properties. Additionally, our results allow for identifying synthetic membranes with a few well defined lipids as optimal plasma membrane mimetics.

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Section: Resultsmentioning

confidence: 99%

Mechanical properties of plasma membrane vesicles correlate with lipid order, viscosity and cell density

et al. 2019

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“…Those authors concluded that dithiothreitol could induce the “unfolded protein response” and this would lead to an over‐expression of the SPT LCB1 subunit mRNA, partially recovering its activity 76 . Dithiothreitol was also shown to affect lipid‐lipid and lipid–protein interactions and to integrate directly into lipid membranes 77 …”

Section: Discussionmentioning

confidence: 99%

CHO/LY‐B cell growth under limiting sphingolipid supply: Correlation between lipid composition and biophysical properties of sphingolipid‐restricted cell membranes

et al. 2021

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“…For example, GPMVs prepared with dithiothreitol (DTT) and paraformaldehyde (PFA) exhibit a demixing temperature about 10-20°C higher than that of GPMVs derived using N-Ethylmaleimide (NEM) (26). DTT has been shown to affect lipidlipid and lipid-protein interactions and to integrate directly into lipid membranes (32,33). To figure out potential differences in mechanical properties, the bending rigidity of GPMVs formed using these two most widely employed methods was assessed.…”

Section: Influence Of Gpmv Isolation Chemicalsmentioning

confidence: 99%

Mechanical properties of plasma membrane vesicles correlate with lipid order and viscosity and depend on cell density

Steinkühler

Sezgin

Urbančič

et al. 2019

Preprint

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Plasma membranes dynamically respond to external cues and changing environment. Quantitative measurements of these adaptations can elucidate the mechanism that cells exploit to survive, adapt and function. However, cell-based assays are affected by active processes while measurements on synthetic models suffer from compositional limitations.Here, as a model system we employ giant plasma membrane vesicles (GPMVs), which largely preserve the plasma membrane lipidome and proteome. From analysis of fluorescence emission and lifetime of environment-sensitive dyes, and membrane shape fluctuations, we investigate how plasma membrane order, viscosity and bending rigidity are affected by different stimuli such as cell seeding density in three different cell models. Our studies reveal that bending rigidity of plasma membranes vary with lipid order and microviscosity in a highly correlated fashion. Thus, readouts from polarity-and viscosity-sensitive probes represent a promising indicator of membrane mechanical properties. Quantitative analysis of the data allows for comparison to synthetic lipid membranes as plasma membrane mimetics.

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Dithiothreitol Raises Transition Temperatures in Giant Plasma Membrane Vesicles

Cited by 4 publications

References 0 publications

Mechanical properties of plasma membrane vesicles correlate with lipid order, viscosity and cell density

Mechanical properties of plasma membrane vesicles correlate with lipid order, viscosity and cell density

CHO/LY‐B cell growth under limiting sphingolipid supply: Correlation between lipid composition and biophysical properties of sphingolipid‐restricted cell membranes

Mechanical properties of plasma membrane vesicles correlate with lipid order and viscosity and depend on cell density

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