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

STAR Protocols

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

[1] Institute of Molecular Medicine, College of Future Technology, Peking University, Beijing 100871, China;

关键词: Cell Membrane;    Microscopy;    Molecular Biology;    Molecular/Chemical Probes;    Protein expression and purification;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: 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).

【 授权许可】

Unknown