Cell-specific mitochondria affinity purification (CS-MAP) from Caenorhabditis elegans

Arnaud Ahier; Tessa Onraet; Steven Zuryn

doi:10.1016/j.xpro.2021.100952

Cell-specific mitochondria affinity purification (CS-MAP) from Caenorhabditis elegans

STAR Protoc. 2021 Nov 16;2(4):100952. doi: 10.1016/j.xpro.2021.100952. eCollection 2021 Dec 17.

Authors

Arnaud Ahier¹, Tessa Onraet¹, Steven Zuryn¹

Affiliation

¹ Clem Jones Centre for Ageing Dementia Research, Queensland Brain Institute, The University of Queensland, Brisbane, QLD 4072, Australia.

Abstract

Cell-Specific Mitochondria Affinity Purification (CS-MAP) enables isolation and purification of intact mitochondria from individual cell types of Caenorhabditis elegans. The approach is based on the cell-specific expression of a recombinant hemagglutinin (HA)-tag fused to the TOMM-20 protein that decorates the surface of mitochondria, thereby allowing their immunomagnetic purification. This protocol describes the CS-MAP procedure performed on large populations of animals. The purified mitochondria are suitable for subsequent nucleic acid, protein, and functional analyses. For complete details on the use and execution of this protocol, please refer to Ahier et al. (2018, 2021).

Keywords: Cell Biology; Cell separation/fractionation; Metabolism; Model Organisms; Molecular Biology; Protein Biochemistry; Protein expression and purification.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Caenorhabditis elegans / cytology*
Cytological Techniques / methods*
Immunologic Techniques / methods*
Mitochondria* / chemistry
Mitochondria* / metabolism
Mitochondria* / physiology

Grants and funding

P40 OD010440/OD/NIH HHS/United States