Journal Article

Toward dynamic structural biology: Two decades of single-molecule Förster resonance energy transfer

Michalet, Xavier, Alhadid, Yazan, Lerner, Eitan, Ingargiola, Antonino, Weiss, Shimon, Cordes, Thorben, Chung, SangYoon

2018

Overview

Structural techniques such as x-ray crystallography and electron microscopy give insight into how macromolecules function by providing snapshots of different conformational states. Function also depends on the path between those states, but to see that path involves watching single molecules move. This became possible with the advent of single-molecule Förster resonance energy transfer (smFRET), which was first implemented in 1996. Lerner et al. review how smFRET has been used to study macromolecules in action, providing mechanistic insights into processes such as DNA repair, transcription, and translation. They also describe current limitations of the approach and suggest how future developments may expand the applications of smFRET. Science , this issue p. eaan1133 Classical structural biology can only provide static snapshots of biomacromolecules. Single-molecule Förster resonance energy transfer (smFRET) paved the way for studying dynamics in macromolecular structures under biologically relevant conditions. Since its first implementation in 1996, smFRET experiments have confirmed previously hypothesized mechanisms and provided new insights into many fundamental biological processes, such as DNA maintenance and repair, transcription, translation, and membrane transport. We review 22 years of contributions of smFRET to our understanding of basic mechanisms in biochemistry, molecular biology, and structural biology. Additionally, building on current state-of-the-art implementations of smFRET, we highlight possible future directions for smFRET in applications such as biosensing, high-throughput screening, and molecular diagnostics.

• Publisher: The American Association for the Advancement of Science
• Subject: Atomic force microscopy / Atomic structure / Biochemistry / Biological activity / Biological evolution / Biological research / Biology / Biosensors / Catalysis / Chemical compounds / Crystallography / Deoxyribonucleic acid / DNA / DNA biosynthesis / DNA repair / DNA topoisomerase / Drug screening / Dynamic structural analysis / Dynamics / Electron microscopy / Energy / Energy transfer / Enzymes / Equilibrium / Experiments / Fluorescence / Fluorescence resonance energy transfer / Fluorescence Resonance Energy Transfer - history / Fluorescence Resonance Energy Transfer - methods / Fluorophores / Folding / Heterogeneity / High-throughput screening / History, 20th Century / History, 21st Century / In vivo methods and tests / Macromolecules / Membrane proteins / Microfluidics / Microscopy / Molecular biology / Molecular Biology - trends / Molecular motors / NMR / Nuclear magnetic resonance / Nuclear reactions / Nuclease / Nucleic Acid Conformation / Nucleic acids / Protein Conformation / Protein folding / Proteins / Repair & maintenance / Replication / Ribonucleic acid / RNA / Screening / Single Molecule Imaging - history / Single Molecule Imaging - methods / Spectroscopy / Subpopulations / Transcription / Translation