Isolation and analysis of fractions enriched in WrappER-associated mitochondria from mouse liver

Nicolò Ilacqua; Irene Anastasia; Luca Pellegrini

doi:10.1016/j.xpro.2021.100752

Isolation and analysis of fractions enriched in WrappER-associated mitochondria from mouse liver

STAR Protoc. 2021 Aug 16;2(3):100752. doi: 10.1016/j.xpro.2021.100752. eCollection 2021 Sep 17.

Authors

Nicolò Ilacqua^{1

2}, Irene Anastasia^{1

2}, Luca Pellegrini^{2

3}

Affiliations

¹ Graduate Program in Neuroscience, Faculty of Medicine, Laval University, Quebec, QC, Canada.
² Mitochondria biology laboratory, Brain Research Center, Quebec, QC, Canada.
³ Dept. of Molecular Biology, Medical Biochemistry, and Pathology, Faculty of Medicine, Laval University, Quebec, QC, Canada.

Abstract

The endoplasmic reticulum (ER) plays a central role in lipid homeostasis, but the role of individual ER subdomains in lipid biology has not been elucidated. WrappER is a curved wrapping type of rough-ER that establishes extensive contacts with almost every mitochondria of the hepatocyte in the mouse liver. Here, we describe a protocol for isolation of fractions enriched in wrappER-associated mitochondria from the mouse liver. We also provide techniques for assessing its quality by electron microscopy and biochemical/proteomic analysis. For complete information on the use and execution of this protocol, please refer to Anastasia et al. (2021).

Keywords: Cell Biology; Cell separation/fractionation; Metabolism; Protein Biochemistry.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Endoplasmic Reticulum*
Liver / chemistry
Liver / cytology*
Mice
Microscopy, Electron
Mitochondria, Liver*
Molecular Biology / instrumentation
Molecular Biology / methods*
Proteomics / methods

Grants and funding

201603PJT-365052/CIHR/Canada