Abstract
In mitochondrial oxidative phosphorylation (Ox-Phos), individual electron transport chain complexes are thought to assemble into supramolecular entities termed supercomplexes (SCs). The technique of blue native (BN) gel electrophoresis has emerged as the method of choice for analyzing SCs. However, the process of sample extraction for BN gel analysis is somewhat tedious and introduces the possibility for experimental artifacts. Here we outline a streamlined method that eliminates a centrifugation step and provides a more representative sampling of a population of mitochondria on the final gel. Using this method, we show that SC composition does not appear to change dynamically with altered mitochondrial function.
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Acknowledgments
Work in the lab of PSB is funded by a grant from the US National Institutes of Health (R01-HL071158). We thank George Porter and Gisela Beutner for critical discussions during the execution of these studies.
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Nadtochiy, S.M., Ngai, M., Brookes, P.S. (2021). Modified Blue Native Gel Approach for Analysis of Respiratory Supercomplexes. In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine . Methods in Molecular Biology, vol 2276. Springer, New York, NY. https://doi.org/10.1007/978-1-0716-1266-8_17
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DOI: https://doi.org/10.1007/978-1-0716-1266-8_17
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