In higher eukaryotes, the mitochondrial electron transport chain consists of five multi-subunit membrane complexes responsible for the generation of cellular ATP. Of these, four complexes are under dual genetic control as they contain subunits encoded by both the mitochondrial and nuclear genomes, thereby adding another layer of complexity to the puzzle of respiratory complex biogenesis. These subunits must be synthesized and assembled in a coordinated manner in order to ensure correct biogenesis of different respiratory complexes. Here, we describe techniques to (1) specifically radiolabel proteins encoded by mtDNA to monitor the rate of synthesis using pulse labeling methods, and (2) analyze the stability, assembly, and turnover of subunits using pulse-chase methods in cultured cells and isolated mitochondria.
|Title of host publication||Mitochondrial DNA|
|Subtitle of host publication||Methods and Protocols|
|Place of Publication||New York NY USA|
|Number of pages||16|
|ISBN (Electronic)||978-1-4939-3040-1 |
|Publication status||Published - 2016|
|Name||Methods in Molecular Biology: Springer Protocols|