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

Clark W.H. Peterson; Kirandeep K. Deol; Milton To; James A. Olzmann

STAR Protocols (Jun 2021)

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

Clark W.H. Peterson,
Kirandeep K. Deol,
Milton To,
James A. Olzmann

Affiliations

Clark W.H. Peterson: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA; Department of Nutritional Sciences and Toxicology, University of California, Berkeley, Berkeley, CA 94720, USA
Kirandeep K. Deol: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA; Department of Nutritional Sciences and Toxicology, University of California, Berkeley, Berkeley, CA 94720, USA
Milton To: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA; Department of Nutritional Sciences and Toxicology, University of California, Berkeley, Berkeley, CA 94720, USA
James A. Olzmann: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA; Department of Nutritional Sciences and Toxicology, University of California, Berkeley, Berkeley, CA 94720, USA; Miller Institute for Basic Research in Science, University of California, Berkeley, Berkeley, CA 94720, USA; Chan Zuckerberg Biohub, San Francisco, CA 94158, USA; Corresponding author

Journal volume & issue: Vol. 2, no. 2
p. 100579

Abstract

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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).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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