Biogenesis of mitochondria. Two-dimensional electrophoretic analysis of mitochondrial translation products in yeast.
1. Mitochondrial translation products of yeast Saccharomyces cerevisiae were separated according to charge as well as molecular weight by a highly resolving two dimensional electorphoretic technique (isoelectric focusing in the first dimension ana SDS-electrophoresis in the second dimension). 2. The major protein components (the oligomeric form of subunit 9 of mitochondrial ATPase, var 1, cytochrome oxidase subunits I, II and III, subunit 6 of mitochondrial ATPase and cytochrome b apoprotein) were identified either from their mobility in SDS-electrophoresis or by using mit- mutants defective in certain mitochondrially made polypeptides. 3. This method allowed the separation of subunit III of cytochrome oxidase and subunit 6 of mitochondrial ATPase which cannot be resolved by conventional SDS-polyacrylamide gel electrophoresis. 4. Subunit II of cytochrome oxiodase resolves in two spots of similar pI values and subunit 6 of mitochondrial ATPase resolves in two spots of similar molecular weight. In both cases the double spots disappear simultaneously following a single mutation in the coresponding structural gene. 5. Total mitochondrial proteins were also resolved two-dimensionally revealing over 100 components. The mitochondrial translation products, with the exception of subunit 9 of mitochondrial ATPase, could be easily recognized among the other mitochondrial proteins.
D000251:Adenosine Triphosphatases / Q000302:isolation & purification; D003576:Electron Transport Complex IV / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005656:Fungal Proteins / Q000096:biosynthesis*; D007525:Isoelectric Focusing; D008928:Mitochondria / Q000378:metabolism*; D014176:Protein Biosynthesis; D012441:Saccharomyces cerevisiae / Q000648:ultrastructure
D005656:Fungal Proteins; D003576:Electron Transport Complex IV; D000251:Adenosine Triphosphatases