Isolation of Plant Mitochondria Using Affinity Purification

Franziska Kuhnert, Andreas P.M. Weber

Research output: Chapter in Book/Conference paperChapterpeer-review

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.

Original languageEnglish
Title of host publicationPlant Mitochondria
Subtitle of host publicationmethods and protocols
EditorsOlivier Van Aken, Allan G. Rasmusson
Place of PublicationUSA
PublisherHumana Press Inc.
Chapter3
Pages25-37
Number of pages13
ISBN (Electronic)978-1-0716-1653-6
ISBN (Print)978-1-0716-1652-9
DOIs
Publication statusPublished - 2022
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume2363
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

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