Subunit analysis of mitochondrial cytochrome c oxidase and cytochrome bc₁ by reversed-phase high-performance liquid chromatography

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• 作者：Peter Kesa^a ; ^b ; ^{peto.kesa@gmail.com" class="auth_mail" title="E-mail the corresponding author} ; Mangesh Bhide^c ; Veronika Lysakova^a ; Andrey Musatov^b ; ^{musatov@saske.sk" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Reversed-phased HPLC ; Cytochrome c oxidase ; Cytochrome bc1 ; Subunits
• 刊名：Analytical Biochemistry
• 出版年：2017
• 出版时间：1 January 2017
• 年：2017
• 卷：516
• 期：Complete
• 页码：6-8
• 全文大小：385 K

文摘

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.