In vitro and in vivo assay of the ER lipid scramblase TMEM41B

Lingzhi Wu; Lu Liu; Bolin Xu; Dong Huang; Xiao-Wei Chen

doi:10.1016/j.xpro.2022.101333

In vitro and in vivo assay of the ER lipid scramblase TMEM41B

STAR Protoc. 2022 Apr 19;3(2):101333. doi: 10.1016/j.xpro.2022.101333. eCollection 2022 Jun 17.

Authors

Lingzhi Wu¹, Lu Liu¹, Bolin Xu¹, Dong Huang¹, Xiao-Wei Chen¹

Affiliation

¹ Institute of Molecular Medicine, College of Future Technology, Peking University, Beijing 100871, China.

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 et al. (2021).

Keywords: Cell Membrane; Microscopy; Molecular Biology; Molecular/Chemical Probes; Protein expression and purification.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biological Transport
Endoplasmic Reticulum* / metabolism
Liposomes / metabolism
Phospholipids* / metabolism

Substances

Liposomes
Phospholipids