First published online April 2, 2004; 10.1104/pp.103.038018
Plant Physiology 134:1450-1459 (2004)
© 2004 American Society of Plant Biologists
BIOENERGETICS AND PHOTOSYNTHESIS
Identification and Characterization of Respirasomes in Potato Mitochondria1
Holger Eubel,
Jesco Heinemeyer and
Hans-Peter Braun*
Institut für Angewandte Genetik, Universität Hannover, D–30419 Hannover, Germany
Plant mitochondria were previously shown to comprise respiratory supercomplexes containing cytochrome c reductase (complex III) and NADH dehydrogenase (complex I) of I1III2 and I2III4 composition. Here we report the discovery of additional supercomplexes in potato (Solanum tuberosum) mitochondria, which are of lower abundance and include cytochrome c oxidase (complex IV). Highly active mitochondria were isolated from potato tubers and stems, solubilized by digitonin, and subsequently analyzed by Blue-native (BN) polyacrylamide gel electrophoresis (PAGE). Visualization of supercomplexes by in-gel activity stains for complex IV revealed five novel supercomplexes of 850, 1,200, 1,850, 2,200, and 3,000 kD in potato tuber mitochondria. These supercomplexes have III2IV1, III2IV2, I1III2IV1, I1III2IV2, and I1III2IV4 compositions as shown by two-dimensional BN/sodium dodecyl sulfate (SDS)-PAGE and BN/BN-PAGE in combination with activity stains for cytochrome c oxidase. Potato stem mitochondria include similar supercomplexes, but complex IV is partially present in a smaller version that lacks the Cox6b protein and possibly other subunits. However, in mitochondria from potato tubers and stems, about 90% of complex IV was present in monomeric form. It was suggested that the I1III2IV4 supercomplex represents a basic unit for respiration in mammalian mitochondria termed respirasome. Respirasomes also occur in potato mitochondria but were of low concentrations under all conditions applied. We speculate that respirasomes are more abundant under in vivo conditions.
1 This work was supported by the Fonds der Chemischen Industrie and the Deutsche Forschungsgemeinschaft (grant BR 1829–7/1.
Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.103.038018.
* Corresponding author; e-mail braun{at}genetik.uni-hannover.de; fax 49511–7623608.
Received December 19, 2003;
returned for revision January 13, 2004;
accepted January 13, 2004.
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