ArticlePloS one2012
GFP's mechanical intermediate states.
Article in PloS one, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Detection of fluorescent protein mechanical switching in cellulo.Cell reports methods · 2024Article
- Electromechanical Photophysics of GFP Packed Inside Viral Protein Cages Probed by Force-Fluorescence Hybrid Single-Molecule Microscopy.Small (Weinheim an der Bergstrasse, Germany) · 2022Article
- Application of FRET Biosensors in Mechanobiology and Mechanopharmacological Screening.Frontiers in bioengineering and biotechnology · 2020Review
- Molecular Tension Sensors: Moving Beyond Force.Current opinion in biomedical engineering · 2019Article
- Tools and Approaches for Dissecting Protein Bacteriocin Import in Gram-Negative Bacteria.Frontiers in microbiology · 2019Review
- High Efficiency Gene Correction in Hematopoietic Cells by Donor-Template-Free CRISPR/Cas9 Genome Editing.Molecular therapy. Nucleic acids · 2018Article
- Mechanically switching single-molecule fluorescence of GFP by unfolding and refolding.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- Titin-Based Nanoparticle Tension Sensors Map High-Magnitude Integrin Forces within Focal Adhesions.Nano letters · 2016Article
- Tissue mechanics and adhesion during embryo development.Developmental biology · 2015Review
- Fluorescence-based force/tension sensors: a novel tool to visualize mechanical forces in structural proteins in live cells.Antioxidants & redox signaling · 2014Review
- Pareto Optimization of Combinatorial Mutagenesis Libraries.IEEE/ACM transactions on computational biology and bioinformaticsArticle
- How to Measure Molecular Forces in Cells: A Guide to Evaluating Genetically-Encoded FRET-Based Tension Sensors.Cellular and molecular bioengineeringArticle
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Green fluorescent protein (GFP) mutants have become the most widely used fluorescence markers in the life sciences, and although they are becoming increasingly popular as mechanical force or strain probes, there is little direct information on how their fluorescence changes when mechanically stretched. Here we derive high-resolution structural models of the mechanical intermediate states of stretched GFP using steered molecular dynamics (SMD) simulations. These structures were used to produce mutants of EGFP and EYFP that mimic GFP's different mechanical intermediates. A spectroscopic analysis revealed that a population of EGFP molecules with a missing N-terminal α-helix was significantly dimmed, while the fluorescence lifetime characteristic of the anionic chromophore state remained unaffected. This suggests a mechanism how N-terminal deletions can switch the protonation state of the chromophore, and how the fluorescence of GFP molecules in response to mechanical disturbance might be turned off.
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