Title: 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,
Abstract: 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.