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

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Endoplasmic reticulum phospholipid scramblase activity revealed after protein reconstitution into giant unilamellar vesicles containing a photostable lipid reporter. / Mathiassen, Patricia P. M.; Menon, Anant K.; Pomorski, Thomas Guenther.

In: Scientific Reports, Vol. 11, No. 1, 14364, 2021.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Mathiassen, PPM, Menon, AK & Pomorski, TG 2021, 'Endoplasmic reticulum phospholipid scramblase activity revealed after protein reconstitution into giant unilamellar vesicles containing a photostable lipid reporter', Scientific Reports, vol. 11, no. 1, 14364. https://doi.org/10.1038/s41598-021-93664-0

APA

Mathiassen, P. P. M., Menon, A. K., & Pomorski, T. G. (2021). Endoplasmic reticulum phospholipid scramblase activity revealed after protein reconstitution into giant unilamellar vesicles containing a photostable lipid reporter. Scientific Reports, 11(1), [14364]. https://doi.org/10.1038/s41598-021-93664-0

Vancouver

Mathiassen PPM, Menon AK, Pomorski TG. Endoplasmic reticulum phospholipid scramblase activity revealed after protein reconstitution into giant unilamellar vesicles containing a photostable lipid reporter. Scientific Reports. 2021;11(1). 14364. https://doi.org/10.1038/s41598-021-93664-0

Author

Mathiassen, Patricia P. M. ; Menon, Anant K. ; Pomorski, Thomas Guenther. / Endoplasmic reticulum phospholipid scramblase activity revealed after protein reconstitution into giant unilamellar vesicles containing a photostable lipid reporter. In: Scientific Reports. 2021 ; Vol. 11, No. 1.

Bibtex

@article{3c64a3a1bef8471f86aa4bcaa78ada09,
title = "Endoplasmic reticulum phospholipid scramblase activity revealed after protein reconstitution into giant unilamellar vesicles containing a photostable lipid reporter",
abstract = "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.",
keywords = "MEMBRANE-PROTEINS, FLIP-FLOP, MECHANISMS, PHOSPHATIDYLSERINE, ELECTROFORMATION, ORGANIZATION, MIXTURES, DYNAMICS, EXPOSURE, MOBILITY",
author = "Mathiassen, {Patricia P. M.} and Menon, {Anant K.} and Pomorski, {Thomas Guenther}",
year = "2021",
doi = "10.1038/s41598-021-93664-0",
language = "English",
volume = "11",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "nature publishing group",
number = "1",

}

RIS

TY - JOUR

T1 - Endoplasmic reticulum phospholipid scramblase activity revealed after protein reconstitution into giant unilamellar vesicles containing a photostable lipid reporter

AU - Mathiassen, Patricia P. M.

AU - Menon, Anant K.

AU - Pomorski, Thomas Guenther

PY - 2021

Y1 - 2021

N2 - 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.

AB - 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.

KW - MEMBRANE-PROTEINS

KW - FLIP-FLOP

KW - MECHANISMS

KW - PHOSPHATIDYLSERINE

KW - ELECTROFORMATION

KW - ORGANIZATION

KW - MIXTURES

KW - DYNAMICS

KW - EXPOSURE

KW - MOBILITY

U2 - 10.1038/s41598-021-93664-0

DO - 10.1038/s41598-021-93664-0

M3 - Journal article

C2 - 34257324

VL - 11

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

IS - 1

M1 - 14364

ER -

ID: 275434626