Assessment of Mitochondrial Protein Composition and Purity by Mass Spectroscopy

Anqiang Tang, Yimin Wang, Nicolas L Taylor

Research output: Chapter in Book/Conference paperChapterpeer-review

Abstract

While the routine mitochondrial extraction and isolation protocols have not fundamentally changed since the introduction of density gradients, the approaches we use to examine the proteome have. The initial characterisation of mitochondrial proteomes was carried out using two-dimensional gel electrophoresis in 2001 and gel spot mass spectrometry have now largely been superseded as the throughput and sensitivity of commercial mass spectrometers increases. Whist many of these early studies established the components of the mitochondrial proteome, as gels were replaced by gel free approaches the numbers of confirmed components rapidly increased. In this chapter we present gel-based approaches for the separation and concentration of mitochondrial proteins for their characterization by mass spectrometry. We also describe two gel-free approaches which can be used to quantity the degree of contamination arising during the isolation of mitochondria. These approaches are equally suitable for studies comparing one treatment to another.

Original languageEnglish
Title of host publicationPlant Mitochondria
Subtitle of host publicationMethods and Protocols
EditorsOliver Van Aken, Allan G. Rasmusson
Place of PublicationUSA
PublisherHumana Press
Chapter11
Pages121-152
Number of pages32
ISBN (Electronic)978-1-0716-1653-6
ISBN (Print)978-1-0716-1652-9
DOIs
Publication statusE-pub ahead of print - 21 Sep 2021

Publication series

NameMethods in molecular biology (Clifton, N.J.)
PublisherHumana Press
ISSN (Print)1064-3745

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