STAR protocols | VOL. 3
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Invitro and in vivo assay of the ER lipid scramblase TMEM41B.

Publication Date Jun 17, 2022

Abstract

Amphipathic phospholipids translocated by scramblases play a central role in facilitating lipid movement across the membrane bilayer, especially at the endoplasmic reticulum (ER) membranes. Here, we present a protocol for assessing the activity of the ER-localized lipid scramblase TMEM41B. We detail an in vitro fluorescent liposome-based phospholipid scrambling assay and in vivo metabolic labeling in living cells using alkyne-choline. The scramblase activity of other VTT (VMP1, TMEM41, and Tvp38) domain-containing proteins, such as TMEM41A and VMP1, can be assayed. For complete details on the use and execution of this protocol, please refer to Huang etal. (2021).

Concepts

Endoplasmic Reticulum Domain-containing Proteins Membrane Bilayer Fluorescent Phospholipid Endoplasmic Reticulum Membranes Reticulum Membranes Endoplasmic Reticulum Lipid Lipid Scramblase Scramblase Activity Lipid Movement

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