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 describe 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.
authors.forename: Colin D Kinz
authors.initials: CDK
authors.lastname: Thompson
authors.forename: Ajeet K
authors.initials: AK
authors.lastname: Sharma
authors.forename: Joachim
authors.initials: J
authors.lastname: Frank
authors.forename: Ruben L
authors.initials: RL
authors.lastname: Gonzalez
authors.forename: Debashish
authors.initials: D
authors.lastname: Chowdhury
mesh_terms: D020285:Cryoelectron Microscopy; D031541:Fluorescence Resonance Energy Transfer; D007700:Kinetics; D008962:Models, Theoretical; D008968:Molecular Conformation; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism; D013816:Thermodynamics