Subunit analysis of mitochondrial cytochrome c oxidase and cytochrome bc by reversed-phase high-performance liquid chromatography.

Anal Biochem

Department of Biophysics, Institute of Experimental Physics, Slovak Academy of Sciences, Watsonova 47, 040 01 Kosice, Slovakia. Electronic address:

Published: January 2017

A rapid separation of the ten nuclearly-encoded subunits of mitochondrial cytochrome c oxidase, and ten out of the eleven subunits of cytochrome bc, was achieved using a short, 50 mm C-reversed-phase column. The short column decreased the elution time 4-7 fold while maintaining the same resolution quality. Elution was similar to a previously published protocol, i.e., a water/acetonitrile elution gradient containing trifluoroacetic acid. Isolated subunits were identified by MALDI-TOF. The rapidity of the described method makes it extremely useful for determining the subunit composition of isolated mitochondrial complexes. The method can be used for both analytical and micro-preparative purposes.

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http://dx.doi.org/10.1016/j.ab.2016.10.006DOI Listing

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