ArticleJournal of magnetic resonance (San Diego, Calif. : 1997)2019
Stability of nitroxide biradical TOTAPOL in biological samples.
Article in Journal of magnetic resonance (San Diego, Calif. : 1997), 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Spatially resolved DNP-assisted NMR illuminates the conformational ensemble of α-synuclein in intact viable cells.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Stability of the polarization agent AsymPolPOK in intact and lysed mammalian cells.Journal of magnetic resonance (San Diego, Calif. : 1997) · 2025Article
- Stability of the polarization agent AsymPolPOK in intact and lysed mammalian cells.bioRxiv : the preprint server for biology · 2024Article
- Expanding the tool box for native structural biology:Science advances · 2024Article
- An EPR Study on Highly Stable Nitroxyl-Nitroxyl Biradicals for Dynamic Nuclear Polarization Applications at High Magnetic Fields.Molecules (Basel, Switzerland) · 2023Article
- A Comparative Study of Nitroxide-Based Biradicals for Dynamic Nuclear Polarization in Cellular Environments.Chembiochem : a European journal of chemical biology · 2022Article
- Highly bioresistant, hydrophilic and rigidly linked trityl-nitroxide biradicals for cellular high-field dynamic nuclear polarization.Chemical science · 2022Article
- Stability of the nitroxide biradical AMUPol in intact and lysed mammalian cells.Journal of magnetic resonance (San Diego, Calif. : 1997) · 2022Article
- TOAC spin-labeled peptides tailored for DNP-NMR studies in lipid membrane environments.Biophysical journal · 2021Article
- Targeted DNP for biomolecular solid-state NMR.Chemical science · 2021Review
- In-Cell NMR of Intact Mammalian Cells Preserved with the Cryoprotectants DMSO and Glycerol Have Similar DNP Performance.Frontiers in molecular biosciences · 2021Article
- Biomolecular Perturbations in In-Cell Dynamic Nuclear Polarization Experiments.Frontiers in molecular biosciences · 2021Article
- In Situ Detection of Endogenous HIV Activation by Dynamic Nuclear Polarization NMR and Flow Cytometry.International journal of molecular sciences · 2020Article
- Biomolecular complex viewed by dynamic nuclear polarization solid-state NMR spectroscopy.Biochemical Society transactions · 2020Review
- Targetable Tetrazine-Based Dynamic Nuclear Polarization Agents for Biological Systems.Chembiochem : a European journal of chemical biology · 2020Article
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4 authors.
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Abstract
We characterize chemical reduction of a nitroxide biradical, TOTAPOL, used in dynamic nuclear polarization (DNP) experiments, specifically probing the stability in whole-cell pellets and lysates, and present a few strategies to stabilize the biradicals for DNP studies. DNP solid-state NMR experiments use paramagnetic species such as nitroxide biradicals to dramatically increase NMR signals. Although there is considerable excitement about using nitroxide-based DNP for detecting the NMR spectra of proteins in whole cells, nitroxide radicals are reduced in minutes in bacterial cell pellets, which we confirm and quantify here. We show that addition of the covalent cysteine blocker N-ethylmaleimide to whole cells significantly slows the rate of reduction, suggesting that cysteine thiol radicals are important to in vivo radical reduction. The use of cell lysates rather than whole cells also slows TOTAPOL reduction, which suggests a possible role for the periplasm and oxidative phosphorylation metabolites in radical degradation. Reduced TOTAPOL in lysates can also be efficiently reoxidized with potassium ferricyanide. These results point to a practical and robust set of strategies for DNP of cellular preparations.
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