Titill: | Sterically shielded spin labels for in-cell EPR spectroscopy: Analysis of stability in reducing environment |
Höfundur: |
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Útgáfa: | 2014-11-20 |
Tungumál: | Enska |
Umfang: | 78-85 |
Háskóli/Stofnun: | Háskóli Íslands University of Iceland |
Svið: | Verkfræði- og náttúruvísindasvið (HÍ) School of Engineering and Natural Sciences (UI) |
Deild: | Raunvísindadeild (HÍ) Faculty of Physical Sciences (UI) Raunvísindastofnun (HÍ) Science Institute (UI) |
Birtist í: | Free Radical Research;49(1) |
ISSN: | 1071-5762 1029-2470 (eISSN) |
DOI: | 10.3109/10715762.2014.979409 |
Efnisorð: | Nitroxide reduction; Aminoxyl radical; Radical stability; Spin labeling; Trityl radical; Litrófsgreining; Efnagreining |
URI: | https://hdl.handle.net/20.500.11815/1954 |
Tilvitnun:A. P. Jagtap, I. Krstic, N. C. Kunjir, R. Hänsel, T. F. Prisner & S. Th. Sigurdsson (2015) Sterically shielded spin labels for in-cell EPR spectroscopy: Analysis of stability in reducing environment, Free Radical Research, 49:1, 78-85DOI: 10.3109/10715762.2014.979409
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Útdráttur:Electron paramagnetic resonance (EPR) spectroscopy is a powerful and widely used technique for studying structure and dynamics of biomolecules under bio-orthogonal conditions. In-cell EPR is an emerging area in this field; however, it is hampered by the reducing environment present in cells, which reduces most nitroxide spin labels to their corresponding diamagnetic N-hydroxyl derivatives. To determine which radicals are best suited for in-cell EPR studies, we systematically studied the effects of substitution on radical stability using five different classes of radicals, specifically piperidine-, imidazolidine-, pyrrolidine-, and isoindoline-based nitroxides as well as the Finland trityl radical. Thermodynamic parameters of nitroxide reduction were determined by cyclic voltammetry; the rate of reduction in the presence of ascorbate, cellular extracts, and after injection into oocytes was measured by continuous-wave EPR spectroscopy. Our study revealed that tetraethyl-substituted nitroxides are good candidates for in-cell EPR studies, in particular pyrrolidine derivatives, which are slightly more stable than the trityl radical.
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Athugasemdir:Post-print (lokagerð höfundar)
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Leyfi:This is an Accepted Manuscript of an article published by Taylor & Francis in Free Radical Research on 20. nov 2014, available online: http://www.tandfonline.com/10.3109/10715762.2014.979409.
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