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Isolation of Plant Mitochondria Using Affinity Purification

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Plant Mitochondria

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2363))

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Abstract

Mitochondria are highly dynamic organelles that are vital for eukaryotic cellular metabolism. Mitochondria account only for a small fraction of the total cell volume. As such, intact organelle isolation is a basic requirement to study mitochondrial biochemistry and physiology. However, established isolation procedures are tedious, and require a substantial amount of tissue material to start with. We addressed this matter and developed a single step purification for Arabidopsis thaliana (Arabidopsis) mitochondria. Ubiquitous expression of an 3xHA-sGFP-TOM5 fusion protein in the outer mitochondrial membrane allows affinity purification by immunocapture in a single step. We here provide a detailed step-by-step protocol for rapid immunocapture of intact and physiologically active mitochondria from Arabidopsis plants.

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Acknowledgments

This work was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)—Project-ID 267205415—SFB 1208 and under Germany’s Excellence Strategy EXC-2048/1, Project ID 390686111.

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Authors and Affiliations

  1. Institute of Plant Biochemistry, Cluster of Excellence on Plant Science (CEPLAS), Heinrich Heine University, Düsseldorf, Germany

    Franziska Kuhnert & Andreas P. M. Weber

Authors
  1. Franziska Kuhnert
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  2. Andreas P. M. Weber
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Corresponding author

Correspondence to Andreas P. M. Weber .

Editor information

Editors and Affiliations

  1. Department of Biology, Lund University, Lund, Sweden

    Olivier Van Aken

  2. Department of Biology, Lund University, Lund, Sweden

    Allan G. Rasmusson

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Kuhnert, F., Weber, A.P.M. (2022). Isolation of Plant Mitochondria Using Affinity Purification. In: Van Aken, O., Rasmusson, A.G. (eds) Plant Mitochondria. Methods in Molecular Biology, vol 2363. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1653-6_3

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  • DOI: https://doi.org/10.1007/978-1-0716-1653-6_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1652-9

  • Online ISBN: 978-1-0716-1653-6

  • eBook Packages: Springer Protocols

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