A splendid molecular factory: De- and reconstruction of the mammalian respiratory chain.

ABSTRACT: Mitochondrial respiratory complexes I to IV and the F₁F_o-ATP synthase (complex V) are large protein assemblies producing the universal cellular energy currency adenosine triphosphate (ATP). Individual complexes have been extensively studied in vitro, but functional co-reconstitution of several mammalian complexes into proteoliposomes, in particular, the combination of a primary pump with the ATP synthase, is less well understood. Here, we present a generic and scalable strategy to purify mammalian respiratory complexes I, III and the ATP synthase from enriched mitochondria in enzymatically fully active form, and procedures to reassemble the complexes into liposomes. A robust functionality can be shown by in situ monitoring of ATP synthesis rates and by using selected

SUBMITTER: Rimle L 

PROVIDER: S-EPMC11962478 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature