Details
| Original language | English |
|---|---|
| Title of host publication | Mitochondrial Function, Part A |
| Subtitle of host publication | Mitochondrial Electron Transport Complexes and Reactive Oxygen Species |
| Editors | William Allison, Immo Scheffler |
| Pages | 183-190 |
| Number of pages | 8 |
| Edition | A |
| Publication status | Published - 5 Apr 2009 |
Publication series
| Name | Methods in Enzymology |
|---|---|
| Number | A |
| Volume | 456 |
| ISSN (Print) | 0076-6879 |
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.
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
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Mitochondrial Function, Part A: Mitochondrial Electron Transport Complexes and Reactive Oxygen Species. ed. / William Allison; Immo Scheffler. A. ed. 2009. p. 183-190 (Methods in Enzymology; Vol. 456, No. A).
Research output: Chapter in book/report/conference proceeding › Contribution to book/anthology › Research › peer review
}
TY - CHAP
T1 - Purification of the Cytochrome c Reductase/Cytochrome c Oxidase Super Complex of Yeast Mitochondria
AU - Braun, Hans Peter
AU - Sunderhaus, Stephanie
AU - Boekema, Egbert J.
AU - Kouřil, Roman
N1 - 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.
PY - 2009/4/5
Y1 - 2009/4/5
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=63449113869&partnerID=8YFLogxK
U2 - 10.1016/S0076-6879(08)04410-8
DO - 10.1016/S0076-6879(08)04410-8
M3 - Contribution to book/anthology
C2 - 19348889
AN - SCOPUS:63449113869
SN - 9780080877761
T3 - Methods in Enzymology
SP - 183
EP - 190
BT - Mitochondrial Function, Part A
A2 - Allison, William
A2 - Scheffler, Immo
ER -