Purification of the Cytochrome c Reductase/Cytochrome c Oxidase Super Complex of Yeast Mitochondria

authored by: Hans Peter Braun, Stephanie Sunderhaus, Egbert J. Boekema, Roman Kouřil
Abstract: The protein complexes of the respiratory chain interact by forming large protein particles called respiratory supercomplexes or "respirasomes". Biochemical characterization of these particles proved to be difficult because of their instability. Here we describe a strategy to isolate and characterize the cytochrome c reductase/cytochrome c oxidase supercomplex of yeast, also termed the III + IV supercomplex, which is based on lactate cultivation of yeast, gentle isolation of mitochondria, membrane solubilization by digitonin, sucrose gradient ultracentrifugation, and native gel electrophoresis. The procedure yields pure forms of two varieties of the III + IV supercomplex composed of dimeric complex III and one or two copies of monomeric complex IV. Supercomplex preparations can be used for physiological or structural investigations.
Organisation(s): Institute of Plant Genetics
External Organisation(s): University of Groningen
Type: Contribution to book/anthology
Pages: 183-190
No. of pages: 8
Publication date: 05.04.2009
Publication status: Published
Peer reviewed: Yes
ASJC Scopus subject areas: Biochemistry, Molecular Biology
Electronic version(s): https://doi.org/10.1016/S0076-6879(08)04410-8 (Access: Unknown)

BibTeX

@inbook{8ad3dc435769430b8000472f07fe5f52,
title = "Purification of the Cytochrome c Reductase/Cytochrome c Oxidase Super Complex of Yeast Mitochondria",
abstract = "The protein complexes of the respiratory chain interact by forming large protein particles called respiratory supercomplexes or {"}respirasomes{"}. Biochemical characterization of these particles proved to be difficult because of their instability. Here we describe a strategy to isolate and characterize the cytochrome c reductase/cytochrome c oxidase supercomplex of yeast, also termed the III + IV supercomplex, which is based on lactate cultivation of yeast, gentle isolation of mitochondria, membrane solubilization by digitonin, sucrose gradient ultracentrifugation, and native gel electrophoresis. The procedure yields pure forms of two varieties of the III + IV supercomplex composed of dimeric complex III and one or two copies of monomeric complex IV. Supercomplex preparations can be used for physiological or structural investigations.",
author = "Braun, {Hans Peter} and Stephanie Sunderhaus and Boekema, {Egbert J.} and Roman Kou{\v r}il",
note = "Funding information: Research in our laboratories was supported by grants of the Deutsche Forschungsgemeinschaft (Br 1829?7/3 and Br 1829?8/1) and the Dutch Science Foundation NWO?CW.",
year = "2009",
month = apr,
day = "5",
doi = "10.1016/S0076-6879(08)04410-8",
language = "English",
isbn = "9780080877761",
series = "Methods in Enzymology",
number = "A",
pages = "183--190",
editor = "William Allison and Immo Scheffler",
booktitle = "Mitochondrial Function, Part A",
edition = "A",
}