Protein dynamics detected by magic-angle spinning relaxation dispersion NMR

Napoli, Federico; Becker, Lea Marie; Schanda, Paul

doi:10.1016/j.sbi.2023.102660

Cited by 4 publications

(3 citation statements)

References 57 publications

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“…13 mg) has become of widespread use (5,6). Akin to solution-state NMR, amide-1 H-detected MAS NMR experiments are employed for many aspects of protein structure and dynamics studies: they are used to obtain backbone resonance assignment (7)(8)(9)(10)(11)(12), determine structures from 1 H N -1 H N distances (6,(13)(14)(15)(16)(17)(18) or longdistance restraints to paramagnetic (14) or 19 F (19) sites, or to characterise protein dynamics (20)(21)(22)(23)(24)(25) (We refer the reader to excellent recent reviews of 1 H-detected MAS NMR and deuteration (26)(27)(28).) Very high MAS frequencies available today with 0.5 mm or 0.7 mm rotors (up to 150 kHz) alleviate the need for deuteration, and protonated proteins are sufficient for structure determination or dynamics studies (29)(30)(31)(32)(33).…”

Section: Introductionmentioning

confidence: 99%

Deuteration of proteins boosted by cell lysates: high-resolution amide and Hα MAS NMR without re-protonation bottleneck

Napoli,

Guan,

Arnaud

et al. 2024

Preprint

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Amide-proton detected magic-angle spinning NMR of deuterated proteins has become a main technique in NMR-based structural biology. In standard deuteration protocols that rely on D2O-based culture media, non-exchangeable amide sites remain deuterated, making these sites unobservable. Here we demonstrate that proteins produced with H2O-based culture medium doped with deuterated cell lysate allow to overcome this “reprotonation bottleneck”, while retaining a high level of deuteration (ca. 80 %) and narrow line widths. We quantified coherence life times of several proteins prepared with this labelling pattern over a range of MAS frequencies (40-100 kHz). We demonstrate that under commonly used conditions (50-60 kHz MAS), amide1H line widths with our labelling approach are comparable to those of perdeuterated proteins and better than those of protonated samples at 100 kHz. For three proteins in the 33-50 kDa size range many previously unobserved amides become visible. We report how to prepare the deuterated cell lysate for our approach from fractions of perdeuterated cultures which are usually discarded, and show that such media can be used identically to commercial media. The residual protonation of Hα sites allows for well-resolved Hα-detected spectra and Hα resonance assignment, exemplified by thede novoassignment of 168 Hα sites in a 39 kDa protein. The approach based on this H2O/cell-lysate deuteration and MAS frequencies compatible with 1.3 or 1.9 mm rotors presents a strong sensitivity benefit over 0.7 mm/100 kHz MAS experiments.

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Section: Introductionmentioning

confidence: 99%

Deuteration of proteins boosted by cell lysates: high-resolution amide and Hα MAS NMR without re-protonation bottleneck

Napoli,

Guan,

Arnaud

et al. 2024

Preprint

Self Cite

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“…Determining the structure and dynamics of proteins and peptides is key to understanding their function in biological systems. Solid-state Nuclear Magnetic Resonance (ssNMR) is a unique technique that can reveal such complex information by probing 13 C, 15 N, 1 H, and 2 H nuclei in biological samples through harnessing the power of magic angle spinning (MAS), cross polarization, proton decoupling, and advanced recoupling techniques. Isotopic labeling, high magnetic fields, and faster spinning rates further increase the sensitivity of ssNMR such that the structure of many proteins and peptides has been determined.…”

Section: Introductionmentioning

confidence: 99%

“…The molecular structure is usually obtained from signal assignments and structural constraints while information on fast molecular motion may be obtained, especially from relaxation. 1,2 Longitudinal relaxation is an incoherent process that returns the longitudinal component of nuclear magnetization to its thermal equilibrium through internal dynamics of the molecule. 3 It has previously been pointed out that spin diffusion may influence the measured relaxation rate constants, 4,5 as the apparent longitudinal relaxation rate constant R * 1 determines the combination of both coherent and incoherent contributions to the change of nuclear magnetization.…”

Section: Introductionmentioning

confidence: 99%

Magic angle spinning effects on longitudinal NMR relaxation: 15N in L-histidine

Afrough,

Christensen,

Jensen

et al. 2023

AIP Advances

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Solid-state magnetic resonance is a unique technique that can reveal the dynamics of complex biological systems with atomic resolution. Longitudinal relaxation is a mechanism that returns longitudinal nuclear magnetization to its thermal equilibrium by incoherent processes. The measured longitudinal relaxation rate constant however represents the combination of both incoherent and coherent contributions to the change of nuclear magnetization. This work demonstrates the effect of magic angle spinning rate on the longitudinal relaxation rate constant in two model compounds: L-histidine hydrochloride monohydrate and glycine serving as proxies for isotopically-enriched biological materials. Most notably, it is demonstrated that the longitudinal N15 relaxation of the two nitrogen nuclei in the imidazole ring in histidine is reduced by almost three orders of magnitude at the condition of rotational resonance with the amine, while the amine relaxation rate constant is increased at these conditions. The observed phenomenon may have radical implications for the solid-state magnetic resonance in biophysics and materials, especially in the proper measurement of dynamics and as a selective serial transfer step in dynamic nuclear polarization.

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Measurement of spin-lattice relaxation times in multiphase polymer systems

Yan,

Zhang

2023

Journal of Magnetic Resonance

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Protein dynamics detected by magic-angle spinning relaxation dispersion NMR

Cited by 4 publications

References 57 publications

Deuteration of proteins boosted by cell lysates: high-resolution amide and Hα MAS NMR without re-protonation bottleneck

Deuteration of proteins boosted by cell lysates: high-resolution amide and Hα MAS NMR without re-protonation bottleneck

Magic angle spinning effects on longitudinal NMR relaxation: 15N in L-histidine

Measurement of spin-lattice relaxation times in multiphase polymer systems

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