Dynamics of giant vesicle assembly from thin lipid films DOI Open Access

Joseph Pazzi,

Anand Bala Subramaniam

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: July 3, 2023

Abstract Giant unilamellar vesicles (GUVs) are micrometer-scale lipid assemblies that emulate key characteristics of biological cell membranes. GUVs can be obtained when solid-supported thin films lipids hydrated in aqueous solutions. However, a comprehensive understanding their assembly dynamics has been lacking, impeding mechanistic insights. Here, we report the time dependence distribution sizes and molar yield through novel ‘stopped-time’ technique. We compare three commonly used techniques, PAPYRUS (Paper-Abetted amPhiphile hYdRation aqUeous Solutions) gentle hydration, electroformation. demonstrate all techniques show sigmoidal curves. Yields increase monotonically before reaching plateau, with surprisingly high yields 60 seconds after hydration. Gentle hydration shows limited evolution contrast to Exploration bud on surfaces uncovers emergence, diameter growth, merging phenomena. To provide explanation our observations, employ thermodynamic budding model. This work expands GUV offers fundamental insights into underlying principles governing this process.

Language: Английский

Preparation of hydrogel-supported giant vesicles via a lipid-coated hydrogel transfer method with electrostatic interaction DOI Creative Commons
Daisuke Saeki,

Kazuo Honma,

Yukihisa Okumura

et al.

Colloids and Surfaces A Physicochemical and Engineering Aspects, Journal Year: 2024, Volume and Issue: unknown, P. 136074 - 136074

Published: Dec. 1, 2024

Language: Английский

Citations

1
Formation of Cell-Sized Liposomes Incorporating a β-Barrel-Structured Porin through Rehydration of a Phospholipid-Membrane Protein Dried Film DOI Creative Commons
Toshiyuki Tosaka, Koki Kamiya

ACS Omega, Journal Year: 2024, Volume and Issue: unknown

Published: Jan. 26, 2024

Giant unilamellar vesicles (GUVs) integrated with membrane proteins (proteo-GUVs) are attractive tools for visualizing protein functions such as enzyme reactions and molecular transportation. In the dehydration–rehydration method, one of methods used to form proteo-GUVs, they formed by using a dried film containing phospholipids through rehydration an alternating current electric field supporting gel. However, these make it difficult proteo-GUVs under physiological salt concentration charged phospholipid conditions or carry risk gel contamination lipid membranes. Therefore, may be harmful proteins. Here, we propose method formation concentrations negatively that do not require To investigate transport modified outer G (OmpG), OmpG-giant OmpG-large (LUVs) were formed. The structure function different mutants reconstituted into LUVs evaluated circular dichroism spectroscopy electrophysiological measurements. addition, OmpG in GUVs was monitoring Ca2+ influx fluorescent molecule leakage from nanopores. We found amount nanopores depended on pore size OmpG. Our forming can applied functional evaluation β-barrel porin biological sensors porin.

Language: Английский

Citations

0
Effect of simulated microgravity on artificial single cell membrane mechanics DOI Creative Commons

Asuwin Prabu R G,

Anagha Manohar,

S. Narendran

et al.

Research Square (Research Square), Journal Year: 2024, Volume and Issue: unknown

Published: May 29, 2024

Abstract The study of cell membrane structures under microgravity is crucial for understanding the inherent physiological and adaptive mechanisms relevant to overcoming challenges in human space travel gaining deeper insight into membrane-protein interactions at reduced gravity. However, dynamics conditions have not unraveled yet. Moreover, complexity cells poses significant when investigating effects on individual components, including membranes. Giant Unilamellar Vesicles (GUVs) serve as valuable cell-mimicking models act artificial cells, providing insights biophysics architecture. Herein, we elucidated simulated conditions. GUVs were synthesized size range 20 ± 2.1 µm examined their morphological changes using a random positioning machine. We observed that well-defined spherical transfigured deformed elongated fluidity increased sevenfold compared normal gravity 48 h. It also noted there reduction microviscosity. shed light mechanics contributes broader responses its implications exploration biomedical applications.

Language: Английский

Citations

0
Assessing the mechanism of facilitated proton transport across GUVs trapped in a microfluidic device DOI Creative Commons

Dominik Ruppelt,

Elena L.M. Ackermann,

Tom Robinson

et al.

Biophysical Journal, Journal Year: 2024, Volume and Issue: 123(18), P. 3267 - 3274

Published: July 26, 2024

Proton transport across lipid membranes is one of the most fundamental reactions that make up living organisms. In vitro, however, study proton can be very challenging due to limitations imposed by concentrations, compartment size, and unstirred layers as well buffer exchange capacity. this study, we have developed a permeation assay based on microfluidic trapping giant vesicles enclosing pH-sensitive dye pyranine address some these challenges. Time-resolved fluorescence imaging upon rapid pH shift enabled us investigate facilitated H+ mediated either channel or carrier. Specifically, compared rates function different gradients gramicidin D carrier carbonyl cyanide-m-chlorophenyl hydrazone. Our results demonstrate efficacy in monitoring distinguishing between channel-like carrier-like mechanism. This groundbreaking result elucidate enigmatic mode mechanism recently discovered natural fibupeptide lugdunin.

Language: Английский

Citations

0
Invagination of Giant Unilamellar Vesicles upon Membrane Mixing with Native Vesicles DOI Creative Commons

Garvita Dhanawat,

Manorama Dey,

Anirudh Singh

et al.

ACS Omega, Journal Year: 2024, Volume and Issue: 9(46), P. 46615 - 46626

Published: Nov. 7, 2024

We demonstrate rapid membrane mixing between GUVs of pure lipid compositions and vesicles (MVs) isolated from the plasma Vero cells, resulting in transfer native lipids proteins to GUVs. The steps involved are docking followed by fusion. show that positively charged essential for docking, components MVs drive interleaflet lateral asymmetry a change tension upon trigger invagination. detected outward inward invagination sites at rim within 10-40 min mixing. extent invaginations depends on cholesterol sphingomyelin (SM) contents Cholesterol content above critical concentration disfavors invaginations, SM is an molecular factor

Language: Английский

Citations

0
Dynamics of giant vesicle assembly from thin lipid films DOI Open Access

Joseph Pazzi,

Anand Bala Subramaniam

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: July 3, 2023

Abstract Giant unilamellar vesicles (GUVs) are micrometer-scale lipid assemblies that emulate key characteristics of biological cell membranes. GUVs can be obtained when solid-supported thin films lipids hydrated in aqueous solutions. However, a comprehensive understanding their assembly dynamics has been lacking, impeding mechanistic insights. Here, we report the time dependence distribution sizes and molar yield through novel ‘stopped-time’ technique. We compare three commonly used techniques, PAPYRUS (Paper-Abetted amPhiphile hYdRation aqUeous Solutions) gentle hydration, electroformation. demonstrate all techniques show sigmoidal curves. Yields increase monotonically before reaching plateau, with surprisingly high yields 60 seconds after hydration. Gentle hydration shows limited evolution contrast to Exploration bud on surfaces uncovers emergence, diameter growth, merging phenomena. To provide explanation our observations, employ thermodynamic budding model. This work expands GUV offers fundamental insights into underlying principles governing this process.

Language: Английский

Citations

0