Different gDIG/gMITO produces specific populations of liquid dispersions that correlates with protein complexes separation on BNGE.
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4gDIG/gMITO can extract all mitochondrial proteins and defines bands that are stable even at higher digitonin concentrations.
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Large liquid dispersions could trap uncomplete solubilized protein aggregates generating so misleading results.
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A specific detergent/protein ratio could set up for each type of mitochondria.
Abstract
The BlueNative page (BNGE) gel has been the reference technique for studying the electron transport chain organization since it was established 20 years ago. Although the migration of supercomplexes has been demonstrated being real, there are still several concerns about its ability to reveal genuine interactions between respiratory complexes. Moreover, the use of different solubilization conditions generates conflicting interpretations. Here, we thoroughly compare the impact of different digitonin concentrations on the liquid dispersions' physical properties and correlate with the respiratory complexes' migration pattern and supercomplexes. Our results demonstrate that digitonin concentration generates liquid dispersions with specific size and variability critical to distinguish between a real association of complexes from being trapped in the same micelle.
