Analyzing conformational changes in single FRET-labeled A1 parts of archaeal A1AO-ATP synthase
ATP synthases utilize a proton motive force to synthesize ATP. In reverse, these membrane-embedded enzymes can also hydrolyze ATP to pump protons over the membrane. To prevent wasteful ATP hydrolysis, distinct control mechanisms exist for ATP synthases in bacteria, archaea, chloroplasts and mitochondria. Single-molecule F\"orster resonance energy transfer (smFRET) demonstrated that the C-terminus of the rotary subunit epsilon in the Escherichia coli enzyme changes its conformation to block ATP hydrolysis. Previously we investigated the related conformational changes of subunit F of the A1AO-ATP synthase from the archaeon Methanosarcina mazei G\"o1. Here, we analyze the lifetimes of fluorescence donor and acceptor dyes to distinguish between smFRET signals for conformational changes and potential artefacts.
Code (0)
등록된 구현이 없습니다.
Similar Papers 제목 키워드 기반
Towards monitoring conformational changes of the GPCR neurotensin receptor 1 by single-molecule FRET
Neurotensin receptor 1 (NTSR1) is a G protein-coupled receptor that is important for signaling in the brain and the gut. Its agonist ligand neurotensin (NTS), a 13-amino-acid peptide, binds with nanomolar affinity from t…
Observing conformations of single FoF1-ATP synthases in a fast anti-Brownian electrokinetic trap
To monitor conformational changes of individual membrane transporters in liposomes in real time, we attach two fluorophores to selected domains of a protein. Sequential distance changes between the dyes are recorded and …
Optimized green fluorescent protein fused to FoF1-ATP synthase for single-molecule FRET using a fast anti-Brownian electrokinetic trap
Observation times of freely diffusing single molecules in solution are limited by the photophysics of the attached fluorescence markers and by a small observation volume in the femtolitre range that is required for a suf…
Resolving dynamics and function of transient states in single enzyme molecules
We used a hybrid fluorescence spectroscopic toolkit to monitor T4 Lysozyme (T4L) in action. By unraveling the kinetic and dynamic interplay of the conformational states, we sought to elucidate the dynamic structural biol…
Decoding Single Molecule Time Traces with Dynamic Disorder
Single molecule time trajectories of biomolecules provide glimpses into complex folding landscapes that are difficult to visualize using conventional ensemble measurements. Recent experiments and theoretical analyses hav…