Journal of Magnetic Resonance - Vol 193 Issue 2

Direct 13C-detection for carbonyl relaxation studies of protein dynamics
Gabriela Pasat, John S. Zintsmaster and Jeffrey W. Peng
We describe a method that uses direct 13C-detection for measuring rotating-frame carbonyl (13CO) relaxation rates to describe protein functional dynamics. Key advantages of method include the following: (i) unique access to 13CO groups that lack a scalar-coupled 15N–1H group; (ii) insensitivity to 15N/1H exchange-broadening that can derail 1H-detected 15N and HNCO methods; (iii) avoidance of artifacts caused by incomplete water suppression. We demonstrate the approach for both backbone and side-chain 13CO groups. Accuracy of the 13C-detected results is supported by their agreement with those obtained from established HNCO-based approaches. Critically, we show that the 13C-detection approach provides access to the 13CO groups of functionally important residues that are invisible via 1H-detected HNCO methods because of exchange-broadening. Hence, the 13C-based method fills gaps inherent in canonical 1H-detected relaxation experiments, and thus provides a novel complementary tool for NMR studies of biomolecular flexibility.

29Si NMR in solid state with CPMG acquisition under MAS
J.W. Wiench, V.S.-Y. Lin and M. Pruski
A remarkable enhancement of sensitivity can be often achieved in 29Si solid-state NMR by applying the well-known Carr–Purcell–Meiboom–Gill (CPMG) train of rotor-synchronized π pulses during the detection of silicon magnetization. Here, several one- and two-dimensional (1D and 2D) techniques are used to demonstrate the capabilities of this approach. Examples include 1D 29Si{X} CPMAS spectra and 2D 29Si{X} HETCOR spectra of mesoporous silicas, zeolites and minerals, where X = 1H or 27Al. Data processing methods, experimental strategies and sensitivity limits are discussed and illustrated by experiments. The mechanisms of transverse dephasing of 29Si nuclei in solids are analyzed. Fast magic angle spinning, at rates between 25 and 40 kHz, is instrumental in achieving the highest sensitivity gain in some of these experiments. In the case of 29Si–29Si double-quantum techniques, CPMG detection can be exploited to measure homonuclear J-couplings.

Solid-state NMR adiabatic TOBSY sequences provide enhanced sensitivity for multidimensional high-resolution magic-angle-spinning 1H MR spectroscopy
Ovidiu C. Andronesi, Dionyssios Mintzopoulos, Jochem Struppe, Peter M. Black and A. Aria Tzika
We propose a solid-state NMR method that maximizes the advantages of high-resolution magic-angle-spinning (HRMAS) applied to intact biopsies when compared to more conventional liquid-state NMR approaches. Theoretical treatment, numerical simulations and experimental results on intact human brain biopsies are presented. Experimentally, it is proven that an optimized adiabatic TOBSY (TOtal through Bond correlation SpectroscopY) solid-state NMR pulse sequence for two-dimensional 1H–1H homonuclear scalar-coupling longitudinal isotropic mixing provides a 20%–50% improvement in signal-to-noise ratio relative to its liquid-state analogue TOCSY (TOtal Correlation SpectroscopY). For this purpose we have refined the symmetry-based 13C TOBSY pulse sequence for 1H MRS use and compared it to MLEV-16 TOCSY sequence. Both sequences were rotor-synchronized and implemented using WURST-8 adiabatic inversion pulses. As discussed theoretically and shown in simulations, the improved magnetization-transfer comes from actively removing residual dipolar couplings from the average Hamiltonian. Importantly, the solid-state NMR techniques are tailored to perform measurements at low temperatures where sample degradation is reduced. This is the first demonstration of such a concept for HRMAS metabolic profiling of disease processes, including cancer, from biopsies requiring reduced sample degradation for further genomic analysis.

Enhanced resolution in proton solid-state NMR with very-fast MAS experiments
Jean-Paul Amoureux, Bingwen Hu and Julien Trébosc
We present a new smooth amplitude-modulated (SAM) method that allows to observe highly resolved 1H spectra in solid-state NMR. The method, which works mainly at fast or ultra-fast MAS speed (νR > 25 kHz) is complementary to previous methods, such as DUMBO, FSLG/PMLG or symmetry-based sequences. The method is very robust and efficient and does not present line-shape distortions or fake peaks. The main limitation of the method is that it requires a modern console with fast electronics that must be able to define the cosine line-shape in a smooth way, without any transient. However, this limitation mainly occurs at ultra-fast MAS where the rotation period is very short.

A frequency-selective REDOR experiment for an SI2 spin system
Eugene Mihaliuk and Terry Gullion
A frequency-selective REDOR experiment is described for SI2 spin systems. The experiment causes the net dipolar dephasing of the S spin to evolve only under the influence of one of the I spins. The experiment is based on a single pair of appropriately phased 90° I-spin pulses, and the I spin causing the S-spin dipolar dephasing is determined by the relative phases between the two 90° pulses. The experiment is demonstrated on a sample of 15N2-l-asparagine.

3D 1H–13C–14N correlation solid-state NMR spectrum
Renée Siegel, Julien Trébosc, Jean-Paul Amoureux, and Zhehong Gan
Nitrogen-14 (spin I = 1) has always been a nucleus difficult to observe in solid-state NMR and until recently its observation was restricted to one-dimensional (1D) spectra. We present here the first 3D 1H–13C–14N NMR correlation spectrum. This spectrum was acquired on a test sample l-histidine·HCl·H2O using a recently developed technique, which consists in indirectly observing 14N nuclei via dipolar recoupling with an HMQC-type experiment.