Quantitative Connection Between Ensemble Thermodynamics and Single-Molecule Kinetics: A Case Study Using Cryogenic Electron Microscopy and Single-Molecule Fluorescence Resonance Energy Transfer Investigations of the Ribosome
  At equilibrium, thermodynamic and kinetic information can be extracted from
biomolecular energy landscapes by many techniques. However, while static,
ensemble techniques yield thermodynamic data, often only dynamic,
single-molecule techniques can yield the kinetic data that describes
transition-state energy barriers. Here we present a generalized framework based
upon dwell-time distributions that can be used to connect such static, ensemble
techniques with dynamic, single-molecule techniques, and thus characterize
energy landscapes to greater resolutions. We demonstrate the utility of this
framework by applying it to cryogenic electron microscopy (cryo-EM) and
single-molecule fluorescence resonance energy transfer (smFRET) studies of the
bacterial ribosomal pre-translocation complex. Among other benefits,
application of this framework to these data explains why two transient,
intermediate conformations of the pre-translocation complex, which are observed
in a cryo-EM study, may not be observed in several smFRET studies.
