title: Subunit analysis of mitochondrial cytochrome c oxidase and cytochrome bc1by reversed-phase high-performance liquid chromatography.
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.
authors.affiliation: Department of Biochemistry, Institute of Chemistry, P. J. Safarik University in Kosice, Moyzesova 11, 040 01 Kosice, Slovakia; Department of Biophysics, Institute of Experimental Physics, Slovak Academy of Sciences, Watsonova 47, 040 01 Kosice, Slovakia. Electronic address: peto.kesa@gmail.com.
authors.forename: Peter
authors.initials: P
authors.lastname: Kesa
authors.affiliation: Department of Microbiology and Immunology, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 04181 Kosice, Slovakia.
authors.forename: Mangesh
authors.initials: M
authors.lastname: Bhide
authors.affiliation: Department of Biochemistry, Institute of Chemistry, P. J. Safarik University in Kosice, Moyzesova 11, 040 01 Kosice, Slovakia.
authors.forename: Veronika
authors.initials: V
authors.lastname: Lysakova
authors.affiliation: Department of Biophysics, Institute of Experimental Physics, Slovak Academy of Sciences, Watsonova 47, 040 01 Kosice, Slovakia. Electronic address: musatov@saske.sk.
authors.forename: Andrey
authors.initials: A
authors.lastname: Musatov
mesh_terms: D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D014450:Electron Transport Complex III / Q000032:analysis*; D003576:Electron Transport Complex IV / Q000032:analysis*; D008929:Mitochondria, Heart / Q000201:enzymology*; D024101:Mitochondrial Proteins / Q000032:analysis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D024101:Mitochondrial Proteins; D003576:Electron Transport Complex IV; D014450:Electron Transport Complex III
keywords: Cytochrome bc(1); Cytochrome c oxidase; Reversed-phased HPLC; Subunits