An automated single-molecule FRET platform for high-content, multiwell plate screening of biomolecular conformations and dynamics

Single-molecule FRET (smFRET) has become a versatile tool for probing the structure and functional dynamics of biomolecular systems, and is extensively used to address questions ranging from biomolecular folding to drug discovery. Confocal smFRET measurements are amongst the widely used smFRET assay...

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Bibliographic Details
Published in:Nature communications 2023-10, Vol.14 (1), p.6511-6511, Article 6511
Main Authors: Hartmann, Andreas, Sreenivasa, Koushik, Schenkel, Mathias, Chamachi, Neharika, Schake, Philipp, Krainer, Georg, Schlierf, Michael
Format: Article
Language:English
Subjects:
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Accuracy
Automation
Biophysics
Data analysis
Datasets
Deoxyribonucleic acid
DNA
Drug discovery
Dynamics
Experiments
Folding
Format
Humanities and Social Sciences
multidisciplinary
Open source software
Parameters
Protein folding
Proteins
Science
Science (multidisciplinary)
Screening
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Summary:Single-molecule FRET (smFRET) has become a versatile tool for probing the structure and functional dynamics of biomolecular systems, and is extensively used to address questions ranging from biomolecular folding to drug discovery. Confocal smFRET measurements are amongst the widely used smFRET assays and are typically performed in a single-well format. Thus, sampling of many experimental parameters is laborious and time consuming. To address this challenge, we extend here the capabilities of confocal smFRET beyond single-well measurements by integrating a multiwell plate functionality to allow for continuous and automated smFRET measurements. We demonstrate the broad applicability of the multiwell plate assay towards DNA hairpin dynamics, protein folding, competitive and cooperative protein–DNA interactions, and drug-discovery, revealing insights that would be very difficult to achieve with conventional single-well format measurements. For the adaptation into existing instrumentations, we provide a detailed guide and open-source acquisition and analysis software. Single-molecule FRET (smFRET) studies often struggle with large parameter spaces. Here, the authors introduce an automated smFRET platform for multiwell plate screening of biomolecular conformations and dynamics.
ISSN:2041-1723
2041-1723
DOI:10.1038/s41467-023-42232-3