Fast acquisition of multi-dimensional spectra in solid-state NMR enabled by ultra-fast MAS

Laage, Ségolène; Sachleben, Joseph R.; Steuernagel, Stefan; Pierattelli, Roberta; Pintacuda, Guido; Emsley, Lyndon

doi:10.1016/j.jmr.2008.10.019

Cited by 113 publications

(97 citation statements)

References 36 publications

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“…This benefit can be enhanced by doping the sample with paramagnetic compounds to reduce the longitudinal relaxation time of the proton [17][18][19]. In this context, it is useful and possible to develop low-power pulse schemes for all time periods of the experiment including cross-polarization [20][21][22] and homonuclear polarization transfer, e.g., DREAM and finite-pulse RFDR to name two arbitrary chosen possibilities [23][24][25][26].…”

Section: Considerations For Fast Mas Experimentsmentioning

confidence: 99%

Spinning proteins, the faster, the better?

Böckmann

Ernst

Meier

2015

Journal of Magnetic Resonance

137

189

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Magic-angle spinning (MAS) is a technique that is a prerequisite for high-resolution solid-state NMR spectroscopy of proteins and other biomolecules. Recently, the 100 kHz limit for the rotation frequency has been broken, arguably making MAS rotors the man-made objects with the highest rotation frequency. This development is expected to have a significant impact on biomolecular NMR as it facilitates proton detection, which allows to partially compensate the loss in overall sensitivity associated with the small sample amounts that fit into MAS rotors with less than 1 mm outer diameter. Under these conditions, the mass-normalized sensitivity of a small rotor becomes much higher than that of larger-volume rotor.2

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Section: Considerations For Fast Mas Experimentsmentioning

confidence: 99%

Spinning proteins, the faster, the better?

Böckmann

Ernst

Meier

2015

Journal of Magnetic Resonance

137

189

View full text Add to dashboard Cite

show abstract

“…The value of enhancement in resolution and sensitivity depends crucially on the experimental linewidths of proton and nitrogen signals (Chevelkov et al 2003). To improve the resolution of proton detected solid-state NMR experiments, fast magic-angle spinning (MAS) (Paulson et al 2003;Zhou et al 2006;Laage et al 2009), application of homonuclear decoupling sequences (Vinogradov et al 1999;Rossum et al 2003;Salager et al 2009), and reduction of the proton homonuclear coupling network by extensive deuteration have been proposed (Chevelkov et al 2006).…”

Section: Introductionmentioning

confidence: 99%

Optimum levels of exchangeable protons in perdeuterated proteins for proton detection in MAS solid-state NMR spectroscopy

et al. 2009

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We present a systematic study of the effect of the level of exchangeable protons on the observed amide proton linewidth obtained in perdeuterated proteins. Decreasing the amount of D 2 O employed in the crystallization buffer from 90 to 0%, we observe a fourfold increase in linewidth for both 1 H and 15 N resonances. At the same time, we find a gradual increase in the signal-to-noise ratio (SNR) for 1 H-15 N correlations in dipolar coupling based experiments for H 2 O concentrations of up to 40%. Beyond 40%, a significant reduction in SNR is observed. Scalarcoupling based 1 H-15 N correlation experiments yield a nearly constant SNR for samples prepared with B30% H 2 O. Samples in which more H 2 O is employed for crystallization show a significantly reduced NMR intensity. Calculation of the SNR by taking into account the reduction in 1 H T 1 in samples containing more protons (SNR per unit time), yields a maximum SNR for samples crystallized using 30 and 40% H 2 O for scalar and dipolar coupling based experiments, respectively. A sensitivity gain of 3.8 is obtained by increasing the H 2 O concentration from 10 to 40% in the CP based experiment, whereas the linewidth only becomes 1.5 times broader. In general, we find that CP is more favorable compared to INEPT based transfer when the number of possible 1 H, 1 H interactions increases. At low levels of deuteration (C60% H 2 O in the crystallization buffer), resonances from rigid residues are broadened beyond detection. All experiments are carried out at MAS frequency of 24 kHz employing perdeuterated samples of the chicken a-spectrin SH3 domain.

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“…However, measuring PREs in solids is a prime example where traditional detection methods have difficulty to provide sufficient sensitivity. While enhanced relaxation caused by a paramagnetic center can be exploited for fast recycling and condensed data collection approaches relying on 13 C detection (23)(24)(25), the addition of paramagnetic dopants is not appropriate for the quantitative measurement of relaxation times as a source of structural and dynamic information. As a result, site-specific PREs in the solid state have only been reported on one small model protein (GB1) by the use of cysteine-containing mutants with paramagnetic tags attached (26,27).…”

mentioning

confidence: 99%

Structure and backbone dynamics of a microcrystalline metalloprotein by solid-state NMR

Knight

Pell

Bertini

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

179

244

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We introduce a new approach to improve structural and dynamical determination of large metalloproteins using solid-state nuclear magnetic resonance (NMR) with 1 H detection under ultrafast magic angle spinning (MAS). The approach is based on the rapid and sensitive acquisition of an extensive set of 15 N and 13 C nuclear relaxation rates. The system on which we demonstrate these methods is the enzyme Cu, Zn superoxide dismutase (SOD), which coordinates a Cu ion available either in Cu þ (diamagnetic) or Cu 2þ (paramagnetic) form. Paramagnetic relaxation enhancements are obtained from the difference in rates measured in the two forms and are employed as structural constraints for the determination of the protein structure. When added to 1 H-1 H distance restraints, they are shown to yield a twofold improvement of the precision of the structure. Site-specific order parameters and timescales of motion are obtained by a Gaussian axial fluctuation (GAF) analysis of the relaxation rates of the diamagnetic molecule, and interpreted in relation to backbone structure and metal binding. Timescales for motion are found to be in the range of the overall correlation time in solution, where internal motions characterized here would not be observable.paramagnetism | nuclear relaxation rates | copper | microcrystal S tructure determination of proteins plays a central role in understanding key events in biology. Although the structure of many proteins can be obtained from single-crystal X-ray diffraction, or by solution-state nuclear magnetic resonance (NMR) spectroscopy, there is nevertheless a range of important substrates for which structures cannot be determined today. These include immobile systems lacking long-range order such as protein aggregates, large complexes and membrane-bound systems.Solid-state NMR has the unique potential to study, with atomic resolution, systems of this nature, and spectacular progress has been made in this area over the last decade (1). There are today a small handful of structures obtained by solid-state NMR, from microcrystalline samples to fibrils and membrane-associated systems (2). Additionally, solid-state NMR is uniquely sensitive to site-specific protein dynamics over a broad range of timescales, and a number of demonstration studies have recently appeared for model proteins (3).The use of perdeuterated proteins has very recently opened the way to highly sensitive proton-detected solid-state experiments (4). Despite early proof-of-principle papers (5), this approach only became popular with the realization that amide sites must be only partially reprotonated (typically 10-30% back exchange) (6-8) to yield well-resolved 1 H spectra. This represented a significant compromise in sensitivity to gain resolution, and effectively made the determination of internuclear distances impractical, with few exceptions (9-11). We have recently shown how this problem can be completely overcome by using 100% reprotonation of exchangeable sites, without loss of resolution, if perdeuteration is combined wit...

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Fast acquisition of multi-dimensional spectra in solid-state NMR enabled by ultra-fast MAS

Cited by 113 publications

References 36 publications

Spinning proteins, the faster, the better?

Spinning proteins, the faster, the better?

Optimum levels of exchangeable protons in perdeuterated proteins for proton detection in MAS solid-state NMR spectroscopy

Structure and backbone dynamics of a microcrystalline metalloprotein by solid-state NMR

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