scholarly journals Endoplasmic reticulum phospholipid scramblase activity revealed after protein reconstitution into giant unilamellar vesicles containing a photostable lipid reporter

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Patricia P. M. Mathiassen ◽  
Anant K. Menon ◽  
Thomas Günther Pomorski
Keyword(s):  
Endoplasmic Reticulum ◽  
Sodium Dithionite ◽  
Metabolic Energy ◽  
Giant Unilamellar Vesicles ◽  
Unilamellar Vesicles ◽  
Phospholipid Scramblase ◽  
Directional Movement ◽  
Lipid Transporters ◽  
Single Vesicle ◽  
Protein Reconstitution

AbstractTransbilayer movement of phospholipids in biological membranes is mediated by a diverse set of lipid transporters. Among them are scramblases that facilitate a rapid bi-directional movement of lipids without metabolic energy input. Here, we established a new fluorescence microscopy-based assay for detecting phospholipid scramblase activity of membrane proteins upon their reconstitution into giant unilamellar vesicles formed from proteoliposomes by electroformation. The assay is based on chemical bleaching of fluorescence of a photostable ATTO-dye labeled phospholipid with the membrane-impermeant reductant sodium dithionite. We demonstrate that this new methodology is suitable for the study of the scramblase activity of the yeast endoplasmic reticulum at single vesicle level.

scholarly journals Endoplasmic reticulum phospholipid scramblase activity revealed after protein reconstitution into giant unilamellar vesicles containing a photostable lipid reporter

2021 ◽  
Author(s):  
Patricia P. M. Mathiassen ◽  
Anant K. Menon ◽  
Thomas Guenther Pomorski
Keyword(s):  
Endoplasmic Reticulum ◽  
Sodium Dithionite ◽  
Metabolic Energy ◽  
Giant Unilamellar Vesicles ◽  
Unilamellar Vesicles ◽  
Phospholipid Scramblase ◽  
Directional Movement ◽  
Lipid Transporters ◽  
Single Vesicle ◽  
Protein Reconstitution

Transbilayer movement of phospholipids in biological membranes is mediated by a diverse set of lipid transporters. Among them are scramblases that facilitate a rapid bi-directional movement of lipids without metabolic energy input. Here, we established a new fluorescence microscopy-based assay for detecting phospholipid scramblase activity of membrane proteins upon their reconstitution into giant unilamellar vesicles formed from proteoliposomes by electroformation. The assay is based on chemical bleaching of fluorescence of a photostable ATTO-dye labeled phospholipid with the membrane-impermeant reductant sodium dithionite. We demonstrate that this new methodology is suitable for the study of the scramblase activity of the yeast endoplasmic reticulum at single vesicle level.

scholarly journals Effect of the Membrane Composition of Giant Unilamellar Vesicles on Their Budding Probability: A Trade-Off between Elasticity and Preferred Area Difference

Life ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 634
Author(s):  
Ylenia Miele ◽  
Gábor Holló ◽  
István Lagzi ◽  
Federico Rossi
Keyword(s):  
Phospholipid Fatty Acid ◽  
Enzymatic Reaction ◽  
Stimuli Responsive ◽  
Membrane Composition ◽  
Giant Unilamellar Vesicles ◽  
Physical Processes ◽  
Unilamellar Vesicles ◽  
Division Process ◽  
The Origin Of Life ◽  
Area Difference

The budding and division of artificial cells engineered from vesicles and droplets have gained much attention in the past few decades due to an increased interest in designing stimuli-responsive synthetic systems. Proper control of the division process is one of the main challenges in the field of synthetic biology and, especially in the context of the origin of life studies, it would be helpful to look for the simplest chemical and physical processes likely at play in prebiotic conditions. Here we show that pH-sensitive giant unilamellar vesicles composed of mixed phospholipid/fatty acid membranes undergo a budding process, internally fuelled by the urea–urease enzymatic reaction, only for a given range of the membrane composition. A gentle interplay between the effects of the membrane composition on the elasticity and the preferred area difference of the bilayer is responsible for the existence of a narrow range of membrane composition yielding a high probability for budding of the vesicles.

Domain Sorting in Giant Unilamellar Vesicles Adsorbed on Glass

Langmuir ◽  
2021 ◽  
Vol 37 (3) ◽  
pp. 1082-1088
Author(s):  
Chiho Kataoka-Hamai ◽  
Kohsaku Kawakami
Keyword(s):  
Giant Unilamellar Vesicles ◽  
Unilamellar Vesicles

scholarly journals Production of giant unilamellar vesicles and encapsulation of lyotropic nematic liquid crystals

Soft Matter ◽  
2021 ◽  
Author(s):  
Peng Bao ◽  
Daniel A. Paterson ◽  
Sally A. Peyman ◽  
J. Cliff Jones ◽  
Jonathan A. T. Sandoe ◽  
...  
Keyword(s):  
Liquid Crystals ◽  
Nematic Liquid Crystals ◽  
Double Emulsion ◽  
Lyotropic Liquid Crystals ◽  
Giant Unilamellar Vesicles ◽  
Unilamellar Vesicles ◽  
Emulsion Droplets ◽  
Double Emulsion Droplets

We describe a modified microfluidic method for making Giant Unilamellar Vesicles (GUVs) via water/octanol-lipid/water double emulsion droplets and encapsulation of nematic lyotropic liquid crystals (LNLCs).

scholarly journals Inter- and Intra-Plane Interactions Control the Existence of Macrodomains in Asymmetric Giant Unilamellar Vesicles

2021 ◽  
Vol 120 (3) ◽  
pp. 147a
Author(s):  
Thais A. Enoki ◽  
Haden L. Scott ◽  
Gerald W. Feigenson ◽  
Frederick A. Heberle
Keyword(s):  
Giant Unilamellar Vesicles ◽  
Unilamellar Vesicles

Ciprofloxacin Encapsulation Into Giant Unilamellar Vesicles: Membrane Binding and Release

2013 ◽  
Vol 102 (2) ◽  
pp. 694-705 ◽  
Author(s):  
Nóra Kaszás ◽  
Tamás Bozó ◽  
Marianna Budai ◽  
Pál Gróf
Keyword(s):  
Membrane Binding ◽  
Giant Unilamellar Vesicles ◽  
Unilamellar Vesicles
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