TY - JOUR

T1 - Isolation and subfractionation of plant mitochondria for proteomic analysis.

AU - Eubel, Holger

AU - Heazlewood, Joshua L.

AU - Millar, A. Harvey

N1 - Copyright: This record is sourced from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

PY - 2007

Y1 - 2007

N2 - Mitochondria carry out a variety of biochemical processes in plant cells. Their primary role is the oxidation of organic acids via the tricarboxylic acid cycle and the synthesis of ATP coupled to the transfer of electrons from reduced NAD+ to O2 via the electron transport chain. However, they also perform many important secondary functions such as synthesis of nucleotides, amino acids, lipids, and vitamins, they contain their own genome and undertake transcription and translation by some unique mechanisms, they actively import proteins and metabolites from the cytosol by a complex set of carriers and membrane channels, they influence programmed cell death of plants, and they respond to cellular signals such as oxidative stress. To understand the full range of mitochondrial functions in plants, the mechanisms that govern their biogenesis, and the way in which mitochondrial activity is perceived by the nucleus requires precise information about the protein components of these organelles. Isolation of mitochondria to identify their proteomes and the changes in these proteomes during development and environmental stress treatments is already under way. In this chapter we provide methods for isolating mitochondria from different plant tissue types, advice on assessing purity and storage of mitochondrial samples, and approaches to fractionate mitochondria to separate their membranes and soluble compartments from each other for proteome analysis.

AB - Mitochondria carry out a variety of biochemical processes in plant cells. Their primary role is the oxidation of organic acids via the tricarboxylic acid cycle and the synthesis of ATP coupled to the transfer of electrons from reduced NAD+ to O2 via the electron transport chain. However, they also perform many important secondary functions such as synthesis of nucleotides, amino acids, lipids, and vitamins, they contain their own genome and undertake transcription and translation by some unique mechanisms, they actively import proteins and metabolites from the cytosol by a complex set of carriers and membrane channels, they influence programmed cell death of plants, and they respond to cellular signals such as oxidative stress. To understand the full range of mitochondrial functions in plants, the mechanisms that govern their biogenesis, and the way in which mitochondrial activity is perceived by the nucleus requires precise information about the protein components of these organelles. Isolation of mitochondria to identify their proteomes and the changes in these proteomes during development and environmental stress treatments is already under way. In this chapter we provide methods for isolating mitochondria from different plant tissue types, advice on assessing purity and storage of mitochondrial samples, and approaches to fractionate mitochondria to separate their membranes and soluble compartments from each other for proteome analysis.

UR - http://www.scopus.com/inward/record.url?scp=39049176896&partnerID=8YFLogxK

M3 - Article

C2 - 17093302

AN - SCOPUS:39049176896

VL - 355

SP - 49

EP - 62

JO - Methods in molecular biology (Clifton, N.J.)

JF - Methods in molecular biology (Clifton, N.J.)

SN - 1064-3745

ER -