FRET-based dynamic structural biology: Challenges, perspectives and an appeal for open-science practices.

Area of Science:

• Biophysics
• Structural Biology
• Biochemistry

Background:

• Single-molecule Förster Resonance Energy Transfer (smFRET) is a key technique for investigating biomolecular structural dynamics.
• Widespread adoption has driven advancements in sample preparation, measurement, data analysis, and documentation.

Purpose of the Study:

• To consolidate and disseminate best practices for quantitative smFRET experiments.
• To address methodological challenges and foster consensus in the field.
• To encourage open science for advancing structural biology.

Main Methods:

• Collaborative multi-lab studies, including blind tests, to assess experimental accuracy and precision.
• Development of standardized procedures and documentation for smFRET experiments.
• Review of the current state-of-the-art in smFRET methodologies.

Main Results:

• Establishment of improved smFRET procedures and documentation, crucial for data archiving (e.g., PDB-Dev).
• Identification of unresolved methodological issues requiring further investigation.
• Compilation of openly available resources for smFRET practitioners.

Conclusions:

• Standardization and collaboration are vital for reliable quantitative structural studies using smFRET.
• Adoption of open science principles will accelerate progress in the field.
• The paper provides recommendations and resources to guide researchers in smFRET analysis.