ArticleProceedings of the National Academy of Sciences of the United States of America2014
Optimized two-color super resolution imaging of Drp1 during mitochondrial fission with a slow-switching Dronpa variant.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 32 papers.
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Who cites it
32 citing papers in PubMed.
- An emerging role of mitochondrial quality control in bone metabolism: from molecular mechanisms to targeted therapeutic interventions.Cellular and molecular life sciences : CMLS · 2025Review
- Insertion of mNeonGreen into the variable domain of DRP-1 permits visualization of functional endogenous protein.microPublication biology · 2025Article
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- Phenoxythiophene sulfonamide compound B355252 protects neuronal cells against glutamate-induced excitotoxicity by attenuating mitochondrial fission and the nuclear translocation of AIF.Experimental and therapeutic medicine · 2021Article
- Review
- GFP fluorescence tagging alters dynamin-related protein 1 oligomerization dynamics and creates disassembly-refractory puncta to mediate mitochondrial fission.Scientific reports · 2020Article
- Encoding quantized fluorescence states with fractal DNA frameworks.Nature communications · 2020Article
- Between life and death: strategies to reduce phototoxicity in super-resolution microscopy.Journal of physics D: Applied physics · 2020Review
- Posttranslational modifications of mitochondrial fission and fusion proteins in cardiac physiology and pathophysiology.American journal of physiology. Cell physiology · 2019Review
- A unique dynamin-related protein is essential for mitochondrial fission in Toxoplasma gondii.PLoS pathogens · 2019Article
- Adrenergic Regulation of Drp1-Driven Mitochondrial Fission in Cardiac Physio-Pathology.Antioxidants (Basel, Switzerland) · 2018Review
- Long-Term Potentiation Requires a Rapid Burst of Dendritic Mitochondrial Fission during Induction.Neuron · 2018Article
- Switchable Fluorophores for Single-Molecule Localization Microscopy.Chemical reviews · 2018Review
- Optimization of sample preparation and green color imaging using the mNeonGreen fluorescent protein in bacterial cells for photoactivated localization microscopy.Scientific reports · 2018Article
- Optimized protocol for combined PALM-dSTORM imaging.Scientific reports · 2018Article
- Insight into the fission mechanism by quantitative characterization of Drp1 protein distribution in the living cell.Scientific reports · 2018Article
- Unraveling the Thousand Word Picture: An Introduction to Super-Resolution Data Analysis.Chemical reviews · 2017Article
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Authors and funding
6 authors.
Funding
Abstract
We studied the single-molecule photo-switching properties of Dronpa, a green photo-switchable fluorescent protein and a popular marker for photoactivated localization microscopy. We found the excitation light photoactivates as well as deactivates Dronpa single molecules, hindering temporal separation and limiting super resolution. To resolve this limitation, we have developed a slow-switching Dronpa variant, rsKame, featuring a V157L amino acid substitution proximal to the chromophore. The increased steric hindrance generated by the substitution reduced the excitation light-induced photoactivation from the dark to fluorescent state. To demonstrate applicability, we paired rsKame with PAmCherry1 in a two-color photoactivated localization microscopy imaging method to observe the inner and outer mitochondrial membrane structures and selectively labeled dynamin related protein 1 (Drp1), responsible for membrane scission during mitochondrial fission. We determined the diameter and length of Drp1 helical rings encircling mitochondria during fission and showed that, whereas their lengths along mitochondria were not significantly changed, their diameters decreased significantly. These results suggest support for the twistase model of Drp1 constriction, with potential loss of subunits at the helical ends.
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