Subunit analysis of mitochondrial cytochrome c oxidase and cytochrome bc1 by reversed-phase high-performance liquid chromatography |
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| Authors: | Peter Kesa Mangesh Bhide Veronika Lysakova Andrey Musatov |
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| Institution: | 1. Department of Biochemistry, Institute of Chemistry, P. J. Safarik University in Kosice, Moyzesova 11, 040 01 Kosice, Slovakia;2. Department of Biophysics, Institute of Experimental Physics, Slovak Academy of Sciences, Watsonova 47, 040 01 Kosice, Slovakia;3. Department of Microbiology and Immunology, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 04181 Kosice, Slovakia |
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| Abstract: | A rapid separation of the ten nuclearly-encoded subunits of mitochondrial cytochrome c oxidase, and ten out of the eleven subunits of cytochrome bc1, was achieved using a short, 50 mm C18-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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| Keywords: | Reversed-phased HPLC Cytochrome c oxidase Cytochrome bc1 Subunits |
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