Optimized protocol for the identification of lipid droplet proteomes using proximity labeling proteomics in cultured human cells

STAR Protocols

James A. Olzmann¹  Kirandeep K. Deol¹  Milton To²  Clark W.H. Peterson² 

[1] Department of Nutritional Sciences and Toxicology, University of California, Berkeley, Berkeley, CA 94720, USA;Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA;

关键词: Cell Biology;    Cell culture;    Cell Membrane;    Cell separation/fractionation;    Cell-based Assays;    Proteomics;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: Lipid droplets are endoplasmic reticulum-derived neutral lipid storage organelles that play critical roles in cellular lipid and energy homeostasis. Here, we present a protocol for the identification of high-confidence lipid droplet proteomes in a cell culture model. This approach overcomes limitations associated with standard biochemical fractionation techniques, employing an engineered ascorbate peroxidase (APEX2) to biotinylate endogenous lipid droplet proteins in living cells for subsequent purification and identification by proteomics.For complete details on the use and execution of this protocol, please refer to Bersuker et al. (2018).

【 授权许可】

Unknown