|Title||How many molecules of mitochondrial complex I are in a cell?|
|Publication Type||Journal Article|
|Year of Publication||2022|
|Authors||Ansari F, Yoval B, Niatsetskaya Z, Ten V, Wittig I, Galkin A|
|Date Published||2022 Mar 05|
Mitochondrial complex I is the only enzyme responsible for oxidation of matrix NADH and regeneration of NAD+ for catabolism. Nuclear and mtDNA mutations, assembly impairments, and enzyme damage are implicated in inherited diseases, ischemia-reperfusion injury, neurodegeneration, and tumorogenesis. Here we introduce a novel method to measure the absolute content of complex I. The method is based on flavin fluorescence scanning of a polyacrylamide gel after separation of complexes by Clear Native electrophoresis. Using mouse primary astrocytes as an example, we calculated an average value of 2.2 × 105 complex I molecules/cell. Our method can be used for accurate quantification of complex I content.
|Alternate Journal||Anal Biochem|