Selective high-resolution DNP-enhanced NMR of biomolecular binding sites

Marín-Montesinos, Ildefonso; Goyard, David; Gillon, Émilie; Renaudet, Olivier; Imberty, Anne; Hediger, Sabine; Paëpe, Gaël De

doi:10.1039/c8sc05696j

Cited by 23 publications

(32 citation statements)

References 42 publications

(65 reference statements)

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“…57,58 The motivation and objectives of the different approaches were diverse, going from optimization of the sample preparation for best sensitivity 59,60 and biocompatibility, [61][62][63] investigation of the bleaching/quenching effect of the paramagnetic tag [64][65][66] under DNP conditions for its potential use as a structural probe, [67][68][69] selection of a particular protein in a complex environment, 62,63,[70][71][72][73][74][75] to the detection of protein-protein interactions, 76 and the development of highresolution localized techniques through selective DNP. 77…”

Section: Targeted Dnp Versus Conventional Dnpmentioning

confidence: 99%

Targeted DNP for biomolecular solid-state NMR

et al. 2021

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Section: Targeted Dnp Versus Conventional Dnpmentioning

confidence: 99%

Targeted DNP for biomolecular solid-state NMR

et al. 2021

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“…These approaches, however, require extensive perturbation of the cellular milieu and are not applicable to the structural studies of endogenous concentrations of cellular components. Alternatively, the design of targeted DNP agents has been explored, with selectivity achieved either through cysteine chemistry or protein–ligand interactions . The applications of these strategies, however, are limited by the reducing cellular environment or the requirement for a known synthetically accessible ligand of the target protein.…”

Section: Figurementioning

confidence: 99%

Targetable Tetrazine‐Based Dynamic Nuclear Polarization Agents for Biological Systems

2020

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Dynamic nuclear polarization (DNP) has shown great promise as a tool to enhance the nuclear magnetic resonance signals of proteins in the cellular environment. As sensitivity increases, the ability to select and efficiently polarize a specific macromolecule over the cellular background has become desirable. Herein, we address this need and present a tetrazine‐based DNP agent that can be targeted selectively to proteins containing the unnatural amino acid (UAA) norbornene‐lysine. This UAA can be introduced efficiently into the cellular milieu by genetic means. Our approach is bio‐orthogonal and easily adaptable to any protein of interest. We illustrate the scope of our methodology and investigate the DNP transfer mechanisms in several biological systems. Our results shed light on the complex polarization‐transfer pathways in targeted DNP and ultimately pave the way to selective DNP‐enhanced NMR spectroscopy in both bacterial and mammalian cells.

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“…The goal of MAS‐DNP is to maximize the sensitivity of solid‐state NMR experiments while maintaining the high resolution of modern solid‐state NMR. Despite several recent demonstrations of the utility of the targeted polarization agents, the mainstream approach to MAS‐DNP remains to uniformly polarize the sample of interest to the highest possible extent in a given time using soluble radicals in the presence of an appropriate solvent that serves as a mediator. The polarization buildup rate and the maximum steady‐state polarization are equally important parameters to optimize in a DNP experiment.…”

Section: The Mas‐dnp Processmentioning

confidence: 99%

Recent Advances in Magic Angle Spinning‐Dynamic Nuclear Polarization Methodology

Kaminker

2019

Israel Journal of Chemistry

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Dynamic nuclear polarization (DNP) is a method to boost the nuclear magnetic resonance (NMR) signal and thus alleviate the main limitation of NMR spectroscopy-its low sensitivity. One particularly successful methodology is the combination of DNP with magic angle spinning (MAS). MAS-DNP enables orders of magnitude signal enhancements in solid-state NMR experiments. The success of modern MAS-DNP stems from a combination of multiple developments in hardware, polarization agents design, sample preparation, theoretical understanding, and experimental methodology. Altogether these developments have allowed numerous breakthrough applications in biology and materials science. Despite its proven successes, ongoing research aims to further optimize hardware and polarization agent chemistry. Recent advances in MAS-DNP methodology that will allow unprecedented sensitivity in novel future applications are reviewed in this manuscript. Figure 3. (a) Schematic (left) and photograph (right) of the spinning module of nitrogen drive and bearing gas/helium-cooled design. (b-c) Schematics of the closed-loop helium-based DNP systems as developed (b) at the University of Grenoble Alpes and (c) at the Osaka University. Adapted from references 53, 48, 49 respectively.

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Selective high-resolution DNP-enhanced NMR of biomolecular binding sites

Abstract: Sel-DNP allows selective recovery of high-resolution information from protein binding sites by DNP-enhanced NMR.

Cited by 23 publications

References 42 publications

Targeted DNP for biomolecular solid-state NMR

Targeted DNP for biomolecular solid-state NMR

Targetable Tetrazine‐Based Dynamic Nuclear Polarization Agents for Biological Systems

Recent Advances in Magic Angle Spinning‐Dynamic Nuclear Polarization Methodology

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