ArticleProceedings of the National Academy of Sciences of the United States of America2017
Mechanically switching single-molecule fluorescence of GFP by unfolding and refolding.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed, 63 citations in OpenAlex.
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- Emergent Chemical Reactivity and Complexity of RNA Condensates.Angewandte Chemie (International ed. in English) · 2025Article
- Hydrogels with prestressed tensegrity structures.Nature communications · 2025Article
- Chemically Tagging Cargo for Specific Packaging inside and on the Surface of Virus-like Particles.ACS nano · 2024Article
- "Forcing" new interpretations of molecular tension sensor studies.Cell reports methods · 2024Article
- Detection of fluorescent protein mechanical switching in cellulo.Cell reports methods · 2024Article
- Protein-Decorated Microbubbles for Ultrasound-Mediated Cell Surface Manipulation.ACS applied bio materials · 2023Article
- Development of Single Molecule Techniques for Sensing and Manipulation of CRISPR and Polymerase Enzymes.Small (Weinheim an der Bergstrasse, Germany) · 2023Review
- Nuclear lamina strain states revealed by intermolecular force biosensor.Nature communications · 2023Article
- Looking at Biomolecular Interactions through the Lens of Correlated Fluorescence Microscopy and Optical Tweezers.International journal of molecular sciences · 2023Review
- Single-molecule mechanical studies of chaperones and their clients.Biophysics reviews · 2022Review
- Optical tweezers across scales in cell biology.Trends in cell biology · 2022Review
- One-step asparaginyl endopeptidase (RSC chemical biology · 2022Article
- Electromechanical Photophysics of GFP Packed Inside Viral Protein Cages Probed by Force-Fluorescence Hybrid Single-Molecule Microscopy.Small (Weinheim an der Bergstrasse, Germany) · 2022Article
- Co-condensation of proteins with single- and double-stranded DNA.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Next generation single-molecule techniques: Imaging, labeling, and manipulation in vitro and in cellulo.Molecular cell · 2022Review
- GEM-IL: A highly responsive fluorescent lactate indicator.Cell reports methods · 2021Article
- Protein nanomechanics in biological context.Biophysical reviews · 2021Review
- Effects of Ligand Binding on the Energy Landscape of Acyl-CoA-Binding Protein.Biophysical journal · 2020Article
- An inkjet-printed polysaccharide matrix for on-chip sample preparation in point-of-care cell counting chambers.RSC advances · 2020Article
Corrections and comments
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Authors and funding
2 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Green fluorescent protein (GFP) variants are widely used as genetically encoded fluorescent fusion tags, and there is an increasing interest in engineering their structure to develop in vivo optical sensors, such as for optogenetics and force transduction. Ensemble experiments have shown that the fluorescence of GFP is quenched upon denaturation. Here we study the dependence of fluorescence on protein structure by driving single molecules of GFP into different conformational states with optical tweezers and simultaneously probing the chromophore with fluorescence. Our results show that fluorescence is lost during the earliest events in unfolding, 3.5 ms before secondary structure is disrupted. No fluorescence is observed from the unfolding intermediates or the ensemble of compact and extended states populated during refolding. We further demonstrate that GFP can be mechanically switched between emissive and dark states. These data definitively establish that complete structural integrity is necessary to observe single-molecule fluorescence of GFP.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.