Exploring the structure and phase behavior of plasma membrane vesicles under extreme environmental conditions

Seeliger, Janine; Erwin, Nelli; Rosin, Christopher D.; Kahse, Marie; Weise, Katrin; Winter, Roland

doi:10.1039/c4cp05845c

Cited by 12 publications

(12 citation statements)

References 39 publications

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“…Another contribution to the low values of bending rigidities of GPMVs might be the high membrane protein content (about 40% by mass is contained in GPMVs 57 ), and proteins and peptides are often observed to reduce membrane bending rigidity 20,58–60 . GPMV membrane softening by transmembrane proteins is also consistent with lower bending rigidity values ( κ ≈ 10 k B T ) found in GPMVs derived from HEK cells overexpressing membrane protein 15 .…”

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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“…Moreover, a possible effect of native-like spatial heterogeneities in the distribution of anionic lipids on the membrane partitioning of K-Ras4B is analyzed by using giant plasma membrane vesicles (GPMVs) that also contain a low amount of PI and a sufficient amount of PIP 2 . 25,26 By using proteincontaining GPMVs and lipid extracts thereof, the influence of membrane proteins on Ras membrane insertion can be evaluated, too. As other studies proposed critical roles for specific lipids, such as PS, in mediating the formation, stability and dynamics of Ras nanoclusters, 15,27 we finally compare the membrane interaction behavior of K-Ras4B in the various model membrane systems analyzed here and in previous experiments 10 to that of N-Ras in neutral one-component lipid bilayers and GPMVs as investigated here and in heterogeneous model membrane systems studied earlier.…”

Section: Introductionmentioning

confidence: 99%

Lipoprotein insertion into membranes of various complexity: lipid sorting, interfacial adsorption and protein clustering

Erwin

Sperlich

Garivet

et al. 2016

Phys. Chem. Chem. Phys.

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In a combined chemical-biological and biophysical approach we explored the membrane partitioning of the lipidated signaling proteins N-Ras and K-Ras4B into membrane systems of different complexity, ranging from one-component lipid bilayers and anionic binary and ternary heterogeneous membrane systems even up to partitioning studies on protein-free and protein-containing giant plasma membrane vesicles (GPMVs). To yield a pictorial view of the localization process, imaging using confocal laser scanning and atomic force microscopy was performed. The results reveal pronounced isoform-specific differences regarding the lateral distribution and formation of protein-rich membrane domains. Line tension is one of the key parameters controlling not only the size and dynamic properties of segregated lipid domains but also the partitioning process of N-Ras that acts as a lineactant. The formation of N-Ras protein clusters is even recorded for almost vanishing hydrophobic mismatch. Conversely, for K-Ras4B, selective localization and clustering are electrostatically mediated by its polybasic farnesylated C-terminus. The formation of K-Ras4B clusters is also observed for the multi-component GPMV membrane, i.e., it seems to be a general phenomenon, largely independent of the details of the membrane composition, including the anionic charge density of lipid headgroups. Our data indicate that unspecific and entropy-driven membrane-mediated interactions play a major role in the partitioning behavior, thus relaxing the need for a multitude of fine-tuned interactions. Such a scenario seems also to be reasonable recalling the high dynamic nature of cellular membranes. Finally, we note that even relatively simple models of heterogeneous membranes are able to reproduce many of the properties of much more complex biological membranes.

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“…A number of vesiculating agents such as DTT/PFA, N‐ethylmaleimide, pyruvic aldehyde, acetaldehyde, glyoxal, pyridoxal, iodoacetate, and p‐chloro‐mercuribenzoate are used for the generation of GPMV. Among them, DTT/PFA is the most widely used vesiculating agent for the generation of GPMV . Here we have also chosen DTT/PFA as a vesiculating agent.…”

Section: Resultsmentioning

confidence: 99%

“…For this purpose, MG‐63 cells were pretreated with an actin destabilizer (cytochalasin D) and cholesterol‐sequestering agent (methyl‐β‐cyclodextrin). After that, GPMV were generated by using a combination of chemical vesiculants, dithiothreitol/paraformaldehyde (DTT/PFA) . The generation of GPMV was monitored by fluorescence microscopy and flow cytometry.…”

Section: Introductionmentioning

confidence: 99%

Osteoblast‐Derived Giant Plasma Membrane Vesicles Induce Osteogenic Differentiation of Human Mesenchymal Stem Cells

et al. 2018

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adopted various strategies to achieve the controlled differentiation of mesenchymal stem cells. Use of chemical cues such as dexamethasone, [2] l-ascorbic acid, [3] bone morphogenetic proteins, [4] and β-glycerol phosphate [5] are well-established practices in this regard. In recent years, understanding of mechanotransduction at the cellular and molecular level allows researchers to exploit various physical cues like flow-induced shear stress [6] and matrix stiffness [7] for osteogenic differentiation of hMSC. Another useful strategy in this regard is the application of electrical cues such as pulse-field and low-frequency electromagnetic field [8] on hMSC to induce osteogenic differentiation. Although the aforesaid strategies have found to be efficient, however, a biomimetic approach toward osteogenic differentiation of hMSC is being sought out over the years for tissue engineering. Use of implants [9] and scaffolds [10] with topographical, structural, and compositional characteristics similar to native bone has so far been the most recognized strategy for biomimetic differentiation of the hMSC. In principle, these approaches rely on the biomimicry of the extracellular components of the bone.In vivo, osteoblasts induce osteogenic differentiation of hMSC through paracrine and juxtacrine signaling. The major signaling pathways involved in these cellular processes are, extracellular regulated kinase (Erk)-activated MAPKinase and β1-Integrin signaling, [11] ephrin type B receptor 4 (EphB4)-ephrinB2 signaling [12,13] and Jagged1-mediated Notch signaling. [14,15] Since this cellular cross-talk involves the ligandreceptor interaction between osteoblasts and MSCs, therefore, from a mechanistic point of view, it is possible that a package derived from osteoblast containing the same set of ligands (both soluble and membrane-bound) can act in a similar manner and induces the osteogenic differentiation in hMSC. So far, this concept has been validated in cancer research where people have proved that microvesicles and exosomes released by cancer cells contain a significant amount of signaling molecules and those exosomes modulate the response of neighboring cells. [16] Recently, it has been shown that exosomes, derived from osteoblast are also capable of inducing osteogenic Exosome-mediated differentiation of human mesenchymal stem cell (hMSC) has opened up a new possibility for the directed osteogenic differentiation of hMSC by the osteoblast-derived vesicles. Here, it is hypothesized that giant plasma membrane vesicles (GPMVs) generated from osteoblast, can direct the osteogenic differentiation of hMSC. GPMVs having different membrane characteristics are generated from osteoblast cells (MG-63) using chemical vesiculants. It is deciphered that there is a treatment-dependent variation in the size and complexity in the GPMVs. It is also showed a variation in lipid composition among the GPMVs. It is demonstrated that GPMVs can be fused with hMSC in vitro and such fusion has no detrimental effect on the viability of hMSC. Furth...

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Exploring the structure and phase behavior of plasma membrane vesicles under extreme environmental conditions

Cited by 12 publications

References 39 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

Lipoprotein insertion into membranes of various complexity: lipid sorting, interfacial adsorption and protein clustering

Osteoblast‐Derived Giant Plasma Membrane Vesicles Induce Osteogenic Differentiation of Human Mesenchymal Stem Cells

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