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

Autor: Clark W.H. Peterson, Kirandeep K. Deol, Milton To, James A. Olzmann
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Zdroj: STAR Protocols, Vol 2, Iss 2, Pp 100579- (2021)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2021.100579
Popis: 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).
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