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

ABSTRACT: 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).

SUBMITTER: Peterson CWH

PROVIDER: S-EPMC8190507 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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

Peterson Clark W H CWH Deol Kirandeep K KK To Milton M Olzmann James A JA

STAR protocols 20210605 2

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 ...[more]

PMID: 34151299

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