ArticleNature protocols2013
Site-directed spin-labeling of nucleotides and the use of in-cell EPR to determine long-range distances in a biologically relevant environment.
Article in Nature protocols, 2013. 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, 66 citations in OpenAlex.
- Exploring the dynamics and structure of PpiB in living Escherichia coli cells using electron paramagnetic resonance spectroscopy.Protein science : a publication of the Protein Society · 2024Article
- Conformational Heterogeneity of β-Barrel Membrane Proteins Observed In Situ Using Orthogonal Spin Labels and Pulsed ESR Spectroscopy.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Synthesis of mono-nitroxides and of bis-nitroxides with varying electronic through-bond communication.Organic & biomolecular chemistry · 2023Article
- Towards Profiling of the G-Quadruplex Targeting Drugs in the Living Human Cells Using NMR Spectroscopy.International journal of molecular sciences · 2021Article
- On the modeling of amplitude-sensitive electron spin resonance (ESR) detection using voltage-controlled oscillator (VCO)-based ESR-on-a-chip detectors.Magnetic resonance (Gottingen, Germany) · 2021Article
- Paramagnetic-iterative relaxation matrix approach: extracting PRE-restraints from NOESY spectra for 3D structure elucidation of biomolecules.Journal of biomolecular NMR · 2019Article
- Article
- Phosphoramidite building blocks with protected nitroxides for the synthesis of spin-labeled DNA and RNA.Beilstein journal of organic chemistry · 2018Article
- Synthesis and Electron Spin Relaxation of Tetracarboxylate Pyrroline Nitroxides.The Journal of organic chemistry · 2017Article
- Experimental demonstration of pH-dependent electrostatic catalysis of radical reactions.Chemical science · 2015Article
- Distance Measurement on an Endogenous Membrane Transporter in E. coli Cells and Native Membranes Using EPR Spectroscopy.Angewandte Chemie (International ed. in English) · 2015Article
- New developments in spin labels for pulsed dipolar EPR.Molecules (Basel, Switzerland) · 2014Review
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Double electron-electron resonance (DEER) is an electron paramagnetic resonance (EPR) technique used to determine distance distributions in the nanometer range between spin labels by measuring their dipole-dipole interactions. Here we describe how in-cell DEER can be applied to spin-labeled DNA sequences to unravel their conformations in living cells by long-range distance measurements in cellula. As EPR detects unpaired electron spins only, diamagnetic molecules provide no background and do not reduce detection sensitivity of the specific signal. Compared with in-cell NMR spectroscopy, low concentrations of spin-labeled molecules can be used owing to the higher sensitivity of EPR per spin. This protocol describes the synthesis of the spin labels, their introduction in DNA strands, the injection of labeled DNA solutions in cells and the performance of in-cell EPR measurements. Completion of the entire protocol takes ~20 d.
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