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

verfasst von

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.

Organisationseinheit(en)

Institut für Pflanzengenetik

Externe Organisation(en)

Reichsuniversität Groningen

Typ

Beitrag in Buch/Sammelwerk

Seiten

183-190

Anzahl der Seiten

8

Publikationsdatum

05.04.2009

Publikationsstatus

Veröffentlicht

Peer-reviewed

Ja

ASJC Scopus Sachgebiete

Biochemie, Molekularbiologie

Elektronische Version(en)

https://doi.org/10.1016/S0076-6879(08)04410-8 (Zugang: Unbekannt)