Abstract

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.

Highlights

  • Regulation of plasma membrane curvature and composition governs essential cellular processes

  • giant plasma membrane vesicles (GPMVs) used for fluctuation analysis were osmotically deflated and exhibited a low membrane tension of about 0.01 μN/m

  • GPMVs, which retain most of the compositional complexity of native Plasma membrane (PM), seem to be compatible with the wide range of biophysical tools developed for synthetic giant unilamellar vesicles[5,7]

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Summary

Introduction

Regulation of plasma membrane curvature and composition governs essential cellular processes. 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. By applying a spectrum of established characterization methods to GPMVs, we compare the readout from the used probes in PM and synthetic membranes and found a correlation of molecular order and viscosity with membrane mechanics across a broad range cell lines and culture conditions

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