ArticleJournal of biomolecular NMR2016
Efficient DNP NMR of membrane proteins: sample preparation protocols, sensitivity, and radical location.
Article in Journal of biomolecular NMR, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers.
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Who cites it
35 citing papers in PubMed, 79 citations in OpenAlex.
- Bimodal Cholesterol for Correlative In-Cell DNP Solid-State NMR and Confocal Microscopy of the Plasma Membrane.Journal of the American Chemical Society · 2026Article
- Solid-State NMR Approaches to Study Protein Structure and Protein-Lipid Interactions.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Structural convergence and membrane interactions of AβCommunications chemistry · 2025Article
- Interface-edited solid-state NMR to study cell interfaces.Communications chemistry · 2025Article
- Sorbitol-Based Glass Matrices Enable Dynamic Nuclear Polarization beyond 200 K.The journal of physical chemistry letters · 2024Article
- Resolution in cryogenic solid state NMR: Challenges and solutions.Protein science : a publication of the Protein Society · 2024Review
- DNP-assisted solid-state NMR enables detection of proteins at nanomolar concentrations in fully protonated cellular milieu.Journal of biomolecular NMR · 2024Article
- Contribution of protein conformational heterogeneity to NMR lineshapes at cryogenic temperatures.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
- Heterotypic Interactions between the 40- and 42-Residue Isoforms of β-Amyloid Peptides on Lipid Bilayer Surfaces.ACS chemical neuroscience · 2023Article
- Biradical Polarizing Agents at High Fields.The journal of physical chemistry. B · 2022Article
- Structure ofBiomacromolecules · 2022Article
- DNP-enhanced solid-state NMR spectroscopy of chromatin polymers.Journal of magnetic resonance open · 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
- Application of DNP-enhanced solid-state NMR to studies of amyloid-β peptide interaction with lipid membranes.Chemistry and physics of lipids · 2021Article
- Targeted DNP for biomolecular solid-state NMR.Chemical science · 2021Review
- Characterization of photonic band resonators for DNP NMR of thin film samples at 7 T magnetic field.Journal of magnetic resonance (San Diego, Calif. : 1997) · 2021Article
- Biomolecular Perturbations in In-Cell Dynamic Nuclear Polarization Experiments.Frontiers in molecular biosciences · 2021Article
- Competing Transfer Pathways in Direct and Indirect Dynamic Nuclear Polarization MAS NMR Experiments on HIV-1 Capsid Assemblies: Implications for Sensitivity and Resolution.Magnetic resonance (Gottingen, Germany) · 2021Article
- Dynamic Nuclear Polarization of Biomembrane Assemblies.Biomolecules · 2020Article
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Authors and funding
5 authors at 2 institutions in 1 country.
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Abstract
Although dynamic nuclear polarization (DNP) has dramatically enhanced solid-state NMR spectral sensitivities of many synthetic materials and some biological macromolecules, recent studies of membrane-protein DNP using exogenously doped paramagnetic radicals as polarizing agents have reported varied and sometimes surprisingly limited enhancement factors. This motivated us to carry out a systematic evaluation of sample preparation protocols for optimizing the sensitivity of DNP NMR spectra of membrane-bound peptides and proteins at cryogenic temperatures of ~110 K. We show that mixing the radical with the membrane by direct titration instead of centrifugation gives a significant boost to DNP enhancement. We quantify the relative sensitivity enhancement between AMUPol and TOTAPOL, two commonly used radicals, and between deuterated and protonated lipid membranes. AMUPol shows ~fourfold higher sensitivity enhancement than TOTAPOL, while deuterated lipid membrane does not give net higher sensitivity for the membrane peptides than protonated membrane. Overall, a ~100 fold enhancement between the microwave-on and microwave-off spectra can be achieved on lipid-rich membranes containing conformationally disordered peptides, and absolute sensitivity gains of 105-160 can be obtained between low-temperature DNP spectra and high-temperature non-DNP spectra. We also measured the paramagnetic relaxation enhancement of lipid signals by TOTAPOL and AMUPol, to determine the depths of these two radicals in the lipid bilayer. Our data indicate a bimodal distribution of both radicals, a surface-bound fraction and a membrane-bound fraction where the nitroxides lie at ~10 Å from the membrane surface. TOTAPOL appears to have a higher membrane-embedded fraction than AMUPol. These results should be useful for membrane-protein solid-state NMR studies under DNP conditions and provide insights into how biradicals interact with phospholipid membranes.
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