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Title: A Proximity Labeling Strategy Provides Insights into the Composition and Dynamics of Lipid Droplet Proteomes

Abstract

Lipid droplet (LD) functions are regulated by a complement of integral and peripheral proteins that associate with the bounding LD phospholipid monolayer. Defining the composition of the LD proteome has remained a challenge due to the presence of contaminating proteins in LD-enriched buoyant fractions. To overcome this limitation, we developed a proximity labeling strategy that exploits LD-targeted APEX2 to biotinylate LD proteins in living cells. Application of this approach to two different cell types identified the vast majority of previously validated LD proteins, excluded common contaminating proteins, and revealed new LD proteins. Moreover, quantitative analysis of LD proteome dynamics uncovered a role for endoplasmic reticulum-associated degradation in controlling the composition of the LD proteome. In conclusion, these data provide an important resource for future LD studies and demonstrate the utility of proximity labeling to study the regulation of LD proteomes.

Authors:
 [1];  [1];  [1];  [2];  [3];  [1];  [1];  [1]
  1. Univ. of California, Berkeley, CA (United States)
  2. Max Planck Institute for Biophysical Chemistry, Gottingen (Germany)
  3. SLAC National Accelerator Lab., Menlo Park, CA (United States); Stanford Univ., Stanford, CA (United States)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1463393
Grant/Contract Number:  
AC02-76SF00515; R00DK095921; R01GM112948; R01CA172667; 16GRNT30870005
Resource Type:
Accepted Manuscript
Journal Name:
Developmental Cell
Additional Journal Information:
Journal Volume: 44; Journal Issue: 1; Journal ID: ISSN 1534-5807
Publisher:
Cell Press - Elsevier
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; lipid droplet; proximity labeling; proteome; endoplasmic reticulum; ERAD; ubiquitin; proteasome; APEX; APEX2; biotinylation

Citation Formats

Bersuker, Kirill, Peterson, Clark W. H., To, Milton, Sahl, Steffen J., Savikhin, Victoria, Grossman, Elizabeth A., Nomura, Daniel K., and Olzmann, James A. A Proximity Labeling Strategy Provides Insights into the Composition and Dynamics of Lipid Droplet Proteomes. United States: N. p., 2017. Web. https://doi.org/10.1016/j.devcel.2017.11.020.
Bersuker, Kirill, Peterson, Clark W. H., To, Milton, Sahl, Steffen J., Savikhin, Victoria, Grossman, Elizabeth A., Nomura, Daniel K., & Olzmann, James A. A Proximity Labeling Strategy Provides Insights into the Composition and Dynamics of Lipid Droplet Proteomes. United States. https://doi.org/10.1016/j.devcel.2017.11.020
Bersuker, Kirill, Peterson, Clark W. H., To, Milton, Sahl, Steffen J., Savikhin, Victoria, Grossman, Elizabeth A., Nomura, Daniel K., and Olzmann, James A. Thu . "A Proximity Labeling Strategy Provides Insights into the Composition and Dynamics of Lipid Droplet Proteomes". United States. https://doi.org/10.1016/j.devcel.2017.11.020. https://www.osti.gov/servlets/purl/1463393.
@article{osti_1463393,
title = {A Proximity Labeling Strategy Provides Insights into the Composition and Dynamics of Lipid Droplet Proteomes},
author = {Bersuker, Kirill and Peterson, Clark W. H. and To, Milton and Sahl, Steffen J. and Savikhin, Victoria and Grossman, Elizabeth A. and Nomura, Daniel K. and Olzmann, James A.},
abstractNote = {Lipid droplet (LD) functions are regulated by a complement of integral and peripheral proteins that associate with the bounding LD phospholipid monolayer. Defining the composition of the LD proteome has remained a challenge due to the presence of contaminating proteins in LD-enriched buoyant fractions. To overcome this limitation, we developed a proximity labeling strategy that exploits LD-targeted APEX2 to biotinylate LD proteins in living cells. Application of this approach to two different cell types identified the vast majority of previously validated LD proteins, excluded common contaminating proteins, and revealed new LD proteins. Moreover, quantitative analysis of LD proteome dynamics uncovered a role for endoplasmic reticulum-associated degradation in controlling the composition of the LD proteome. In conclusion, these data provide an important resource for future LD studies and demonstrate the utility of proximity labeling to study the regulation of LD proteomes.},
doi = {10.1016/j.devcel.2017.11.020},
journal = {Developmental Cell},
number = 1,
volume = 44,
place = {United States},
year = {2017},
month = {12}
}

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