Format

Send to

Choose Destination
Methods Mol Biol. 2019;2008:57-72. doi: 10.1007/978-1-4939-9537-0_5.

Identification of Lipid Droplet Proteomes by Proximity Labeling Proteomics Using APEX2.

Author information

1
Department of Nutritional Sciences and Toxicology, University of California, Berkeley, CA, USA.
2
Department of Nutritional Sciences and Toxicology, University of California, Berkeley, CA, USA. olzmann@berkeley.edu.
3
Chan Zuckerberg Biohub, San Francisco, CA, USA. olzmann@berkeley.edu.

Abstract

Lipid droplets (LDs) are ubiquitous lipid storage organelles composed of a neutral lipid core surrounded by a phospholipid monolayer that is decorated with integral and peripheral proteins. Accurate identification of LD proteins using biochemical fractionation methods has been challenging due to the presence of contaminant proteins from co-fractionating organelles. Here, we describe a method to identify high-confidence LD proteomes that employs an engineered ascorbate peroxidase (APEX2) to induce spatially and temporally restricted biotinylation of LD proteins. This proximity labeling method can be broadly applied to define the composition of the LD proteome in any cultured cell line and can be utilized to examine LD proteome dynamics.

KEYWORDS:

APEX; APEX2; Biotinylation; Lipid droplet; Organelle; Proteome; Proximity labeling

PMID:
31124088
PMCID:
PMC6609093
[Available on 2020-01-01]
DOI:
10.1007/978-1-4939-9537-0_5

Supplemental Content

Full text links

Icon for Springer
Loading ...
Support Center