Rotary catalysis of bovine mitochondrial F₁-ATPase studied by single-molecule experiments.

ABSTRACT: The reaction scheme of rotary catalysis and the torque generation mechanism of bovine mitochondrial F₁ (bMF₁) were studied in single-molecule experiments. Under ATP-saturated concentrations, high-speed imaging of a single 40-nm gold bead attached to the γ subunit of bMF₁ showed 2 types of intervening pauses during the rotation that were discriminated by short dwell and long dwell. Using ATPγS as a slowly hydrolyzing ATP derivative as well as using a functional mutant βE188D with slowed ATP hydrolysis, the 2 pausing events were distinctively identified. Buffer-exchange experiments with a nonhydrolyzable analog (AMP-PNP) revealed that the long dwell corresponds to the catalytic dwell, that is, the waiting state for hydrolysis, while it remains elusive which c

SUBMITTER: Kobayashi R 

PROVIDER: S-EPMC6983367 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature