Subunit structures of purified beef mitochondrial cytochrome bc1 complex from liver and heart.
The existence of tissue-specific isozymes of cytochrome c oxidase has been widely documented. We have now studied if there are differences between subunits of mitochondrial bc1 complexes isolated from liver and heart. For this purpose, we have developed a method for the purification of an active ubiquinol-cytochrome c oxidoreductase from adult bovine liver that includes solubilization of submitochondrial particles with deoxycholate, ammonium acetate fractionation, resolubilization with dodecyl maltoside, and ion exchange chromatography. The electrophoretic pattern of the liver preparation showed the presence of 11 subunits, with apparent molecular weights identical to the ones reported for the heart complex. Western blot analysis and isoelectric focusing followed by two-dimensional gels of bc1 complexes from liver and heart were compared, and no qualitative differences were observed. In addition, the high-molecular-weight subunits of the purified complexes from both tissues, subunits I, II, V, and VI, were isolated by PAGE in the presence of Coomasie Blue and subjected to limited proteolysis and to chemical digestion with cyanogen bromide and BNPS-skatol, and the peptide patterns were compared. Finally, two of the small-molecular-weight subunits from the liver complex were isolated (subunits VII and X), partially analyzed by amino terminal sequencing, and found to be identical with the reported sequence of their heart counterparts. The data suggest that, in contrast to the case of cytochrome c oxidase, bc1 complexes from liver and heart do not exhibit tissue-specific differences.
Vázquez-Acevedo
M
M
Departamento de Bioenergética, Universidad Nacional Autónoma de México, México, D.F.
Antaramian
A
A
Corona
N
N
González-Halphen
D
D
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002458:Cell Fractionation; D002852:Chromatography, Ion Exchange; D014450:Electron Transport Complex III / Q000737:chemistry* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000737:chemistry* / Q000302:isolation & purification; D008929:Mitochondria, Heart / Q000201:enzymology*; D008930:Mitochondria, Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009928:Organ Specificity; D011487:Protein Conformation
D007527:Isoenzymes; D014450:Electron Transport Complex III