Triple resonance, triple-quantum NMR studies of dipolar coupled spin systems

Multiple quantum-single quantum correlation experiments are employed for spectral simplification and determination of the relative signs of the couplings. In this study, we have demonstrated the excitation of three nuclei, triple quantum coherences and discussed the information obtainable from such experiments. The experiments have been carried out on doubly labeled acetonitrile and fluoroacetonitrile aligned in liquid crystalline media. The experiment is advantageous in providing many spectral parameters from a single experiment. The coherence pathways involved in the pulse sequence are described using product operators.     

Relaxation-induced dipolar exchange with recoupling-An MAS NMR method for determining heteronuclear distances without irradiating the second spin

A new magic-angle spinning NMR method for distance determination between unlike spins, where one of the two spins in question is not irradiated at all, is introduced. Relaxation-induced dipolar exchange with recoupling (RIDER) experiments can be performed with conventional double-resonance equipment and utilize the familiar π-pulse trains to recouple the heteronuclear dipolar interaction under magic-angle spinning conditions. Longitudinal relaxation of the passive spin during a delay between two recoupling periods results in a dephasing of the heteronuclear coherence and consequently a dephasing of the magnetization detected after the second recoupling period. The information about the dipolar coupling is obtained by recording normalized dephasing curves in a fashion similar to the REDOR experiment. At intermediate mixing times, the dephasing curves also depend on the relaxation properties of the passive spin, i.e., on single- and double-quantum longitudinal relaxation times for the case of I = 1 nuclei, and these relaxation times can be estimated with this new method. To a good approximation, the experiment does not depend on possible quadrupolar interactions of the passive spin, which makes RIDER an attractive method when distances to quadrupolar nuclei are to be determined. The new method is demonstrated experimentally with ¹⁴N and ²H as heteronuclei and observation of ¹³C in natural abundance.     

Heteronuclear dipolar recoupling of half-integer quadrupole nuclei under fast magic angle spinning

An experimental method for the heteronuclear dipolar recoupling of half-integer quadrupole nuclei is proposed. The idea is to manipulate the central transition based on the recoupling technique of spin-polarization-inversion rotary resonance. This method allows the extraction of structural parameters under fast magic-angle spinning. Its validity has been examined by the average Hamiltonian theory and numerical simulations. The initial rotational-echo dephasing arising from the dipolar evolution can be approximated by a parabolic function, from which the heteronuclear van Vleck second moment can be estimated. A factor, estimated from two-spin simulations, is required to account for the effects of the quadrupolar coupling and is rather independent of the geometry and the orders of the spin systems. Our method can facilitate the structural characterization of materials containing half-integer quadrupole nuclei under high-resolution condition. Experimental verification has been carried out on two aluminophosphate systems, namely, AlPO₄-5 and AlPO₄-11.     

Time displacement rotational echo double resonance: heteronuclear dipolar recoupling with suppression of homonuclear interaction under fast magic-angle spinning

We have developed a novel variant of REDOR which is applicable to multiple-spin systems without proton decoupling. The pulse sequence is constructed based on a systematic time displacement of the pi pulses of the conventional REDOR sequence. This so-called time displacement REDOR (td-REDOR) is insensitive to the effect of homonuclear dipole-dipole interaction when the higher order effects are negligible. The validity of td-REDOR has been verified experimentally by the P-31{C-13} measurements on glyphosate at a spinning frequency of 25 kHz. The experimental dephasing curve is in favorable agreement with the simulation data without considering the homonuclear dipole-dipole interactions.     

Dephasing of spin echoes by multiple heteronuclear dipolar interactions in rotational echo double resonance NMR experiments

The application of rotational echo double resonance (REDOR) nuclear magnetic resonance (NMR) for accurate distance measurements has thus far been largely restricted to isolated heteronuclear two-spin systems. In the present paper, the informational content of REDOR curves is explored for systems characterized by multi-spin interactions. To this end, numerical REDOR simulations are presented for cases in which single observe spins S are dipolarly coupled to groups of spins I in distinct geometries. To develop the utility of REDOR for characterizing dipolar couplings in unknown and/or ill-defined geometries, the validity ranges and systematic errors of certain analytical approximations are studied. In the limit of short dipolar evolution times where 0 < deltaS/S0 < or = 0.2 to 0.3, the REDOR difference signal intensity increases approximately proportional to the square of the dipolar evolution time. Here, the curvature depends simply on the second moment M2 characterizing the overall strength of the heterodipolar coupling, irrespective of specific molecular geometries. Fitting experimental REDOR data in this manner produces slight systematic underestimates of M2. However, these errors tend to be counterbalanced by additional systematic errors made by neglecting weak couplings to more remote spins and distribution effects caused by disorder. Based on these findings, the results suggest a convenient method of obtaining site-resolved second moment information in disordered materials.     

Residual 31P, 35,37Cl dipolar coupling in 31P MAS spectra of chlorocyclophosphazenes

In ³¹P MAS NMR spectra of chlorocyclophosphazenes, characteristic splittings have been observed for PCl or PCl₂ groups. At different applied magnetic fields, the fine structure and total width of the patterns change in a characteristic way, demonstrating that the splittings are due to indirect spin–spin and residual dipolar interactions with the chlorine nuclei directly bonded to phosphorus. For trans-nongeminal N₃P₃Cl₃(NMe₂)₃ and N₃P₃Cl₆ as examples, the spectra have been analyzed to obtain information on chlorine nuclear quadrupole coupling constants and ^35,37Cl, ³¹P indirect spin–spin coupling constants. Neglect of these interactions may result in misinterpretations of the multiplicity in ³¹P MAS spectra of chlorophosphazenes.     

REDCAT: a residual dipolar coupling analysis tool

Recent advancements in the utilization of residual dipolar couplings (RDCs) as a means of structure validation and elucidation have demonstrated the need for, not only a more user friendly, but also a more powerful RDC analysis tool. In this paper, we introduce a software package named REsidual Dipolar Coupling Analysis Tool (REDCAT) designed to address the above issues. REDCAT is a user-friendly program with its graphical-user-interface developed in Tcl/Tk, which is highly portable. Furthermore, the computational engine behind this GUI is written in C/C++ and its computational performance is therefore excellent. The modular implementation of REDCAT's algorithms, with separation of the computational engine from the graphical engine allows for flexible and easy command line interaction. This feature can be utilized for the design of automated data analysis sessions. Furthermore, this software package is portable to Linux clusters for high throughput applications. In addition to basic utilities to solve for order tensors and back calculate couplings from a given order tensor and proposed structure, a number of improved algorithms have been incorporated. These include the proper sampling of the Null-space (when the system of linear equations is under-determined), more sophisticated filters for invalid order-tensor identification, error analysis for the identification of the problematic measurements and simulation of the effects of dynamic averaging processes.     

Measurement of one-bond heteronuclear dipolar coupling contributions for amine and diastereotopic methylene protons

One-bond heteronuclear and two-bond homonuclear residual dipolar couplings measured at methylene or amine sites can be utilized as long-range constraints in structure determination of molecules as well as to facilitate characterization of local conformation by stereospecific assignment of diastereotopic protons. We present two J-modulated HMQC type experiments to measure the one-bond heteronuclear dipolar coupling contributions of geminal protons individually. In addition two-bond homonuclear residual dipolar couplings between the diastereotopic protons are also obtained.     

A high sensitivity 3D experiment for measuring Calpha-Halpha residual dipolar coupling constants

A new sensitivity improved approach is presented to measure the Calpha-Halpha scalar and dipolar coupling constants in 13C/15N-labeled proteins using a HA(CA)CONH scheme. The proposed experiment has significantly higher sensitivity than the previously published (HA)CA(CO)NH sequence, and provides accurate and straightforward measurements of the scalar and residual dipolar coupling constants. The sequence is easy to implement, and has been demonstrated on the C-terminal domain of the human Ku-80 protein (152 amino acid residues). On average, sensitivity is improved by 40% for both isotropic and anisotropic samples. The sensitivity enhancement is more pronounced for structured regions than unstructured regions, with an average of 50-60% enhancement being observed in the well-structured regions of the protein.     

Proton-detected scalar coupling based assignment strategies in MAS solid-state NMR spectroscopy applied to perdeuterated proteins

Assignment of proteins in MAS (magic angle spinning) solid-state NMR relies so far on correlations among heteronuclei. This strategy is based on well dispersed resonances in the ¹⁵N dimension. In many complex cases like membrane proteins or amyloid fibrils, an additional frequency dimension is desirable in order to spread the amide resonances. We show here that proton detected HNCO, HNCA, and HNCACB type experiments can successfully be implemented in the solid-state. Coherences are sufficiently long lived to allow pulse schemes of a duration greater than 70 ms before incrementation of the first indirect dimension. The achieved resolution is comparable to the resolution obtained in solution-state NMR experiments. We demonstrate the experiments using a triply labeled sample of the SH3 domain of chicken α-spectrin, which was re-crystallized in H₂O/D₂O using a ratio of 1/9. We employ paramagnetic relaxation enhancement (PRE) using EDTA chelated Cu^II to enable rapid data acquisition.     

Double-quantum filtered rotational-echo double resonance

The homonuclear scalar coupling of a directly bonded 13C-13C pair has been used to create a double-quantum filter (DQF) to remove the natural-abundance 13C background in 13C{15N} rotational-echo double-resonance (REDOR) experiments. The DQF scalar and REDOR dipolar evolution periods are coincident which is important for sensitivity in the event of weak 13C-15N dipolar coupling. Calculated and observed 13C{15N} DQF-REDOR dephasings were in agreement for a test sample of mixed recrystallized labeled alanines. Glycine metabolism in a single uniform-15N soybean leaf labeled for 6 min by 13CO2 was measured quantitatively by 13C{15N} DQF-REDOR with no background interferences.     

Simultaneous extraction of multiple orientational constraints of membrane proteins by 13C-detected N-H dipolar couplings under magic angle spinning

A 13C-detected N-H dipolar coupling technique is introduced for uniaxially mobile membrane proteins for orientation determination using unoriented samples. For proteins undergoing rigid-body uniaxial rotation in the lipid bilayer, the intrinsic equality between the dipolar coupling constants measured in unoriented samples and the anisotropic coupling measured in static oriented samples has been shown recently. Here, we demonstrate that the orientation-sensitive backbone N-H dipolar couplings can be measured with 13C detection using 2D and 3D MAS correlation experiments, so that maximal site resolution can be achieved and multiple orientational constraints can be extracted from each experiment. We demonstrate this technique on the M2 transmembrane peptide of the influenza A virus, where the N-H dipolar couplings of various residues fit to a dipolar wave for a helical tilt angle of 37 degrees , in excellent agreement with data obtained from singly 15N-labeled samples.     

Efficiency at high spinning frequencies of heteronuclear decoupling methods designed to quench rotary resonance

The performance of two recently developed heteronuclear decoupling schemes designed to quench rotary resonance, phase-inverted supercycled sequence for attenuation of rotary resonance (PISSARRO) and high-phase two-pulse phase modulation (high-phase TPPM), are probed at high spinning frequencies. High-phase TPPM may be useful at the n=1 rotary resonance condition while PISSARRO permits efficient decoupling over a broad commonly used range of rf amplitudes, even at very high spinning frequencies. New insights into the response of spin systems to both decoupling schemes have been gained. High-phase TPPM is sensitive to the offsets of remote protons, their chemical shift anisotropies, and the relative orientations of the heteronuclear dipolar and proton chemical shift tensors. Since PISSARRO is virtually immune against such effects, the method is especially suited for very high magnetic fields.     

Multiple resonance heteronuclear decoupling under MAS: dramatic increase of spectral resolution at moderate magnetic field and MAS frequencies

The effects of multiple-resonance heteronuclear decoupling under magic angle spinning (MAS) on the resolution of one-dimensional ¹⁹F and ³¹P and various two-dimensional MAS NMR spectra and on the residual non-refocusable coherence lifetimes in fluorinated aluminophosphate AlPO₄-CJ2, i.e. a compound that contains numerous highly abundant nuclei but no homonuclear spin bath, has been investigated. The design of the four-channel (¹H, ¹⁹F, ²⁷Al, ³¹P) MAS probe used for this study is first described. ¹H and ¹H–²⁷Al double-resonance decouplings allows lengthening the optimized transverse relaxation and increasing the resolution in the ¹⁹F and ³¹P dimensions. Under the application of multi-nuclear decoupling, a two-dimensional ¹⁹F–³¹P CP-HETCOR correlation spectrum for AlPO₄-CJ2 is recorded with unprecedented high-resolution in the two dimensions. Moreover, because ¹H-decoupling increases the ¹⁹F , it has been applied during the entire duration of the 2D NMR experiments, allowing the direct use of residual small interactions to generate ¹⁹F–¹⁹F and ¹⁹F–²⁷Al 2D NMR correlation spectra in AlPO₄-CJ2.     

Double resonance experiments in a single resonance probe: detecting Na–V dipolar interactions in sodium vanadates

²³Na-{⁵¹V} double resonance TRAPDOR experiments are presented on two different sodium vanadates. This is the first time that the heteronuclear dipolar interaction between nuclei, whose Larmor frequencies lie within a range of 0 to 3 MHz is detected in order to monitor connectivity and internuclear distance information in these systems.     

MAS NMR with and without double-quantum filtration at and near the n=0 rotational resonance condition

Spectral lineshapes of MAS NMR spectra of dipolar (re)coupled spin pairs exhibiting considerable chemical shielding anisotropies at and near the so-called n=0 rotational resonance (R2) condition are considered. The n=0 R2 condition is found to be not extremely sharp. Anisotropic interaction parameters such as chemical shielding tensor orientations and the magnitude of the dipolar coupling constant remain sensitively encoded in such lineshapes even when differences in isotropic chemical shielding values of up to 400 Hz (corresponding to ca. half the size of the dipolar coupling constant) are present. Additional double-quantum filtration (DQF) may enhance the sensitivity of spectral lineshapes to anisotropic interaction parameters for even larger differences in isotropic chemical shielding values. The dependence of the DQF efficiency on spin-system parameters as well as on external parameters (Larmor and MAS frequencies) is investigated. Away from R2 conditions a trend to lower DQF efficiencies is found whereas some spin-system parameters are more sensitively encoded in the corresponding spectral lineshapes. Our study is based on numerical simulations, with the known parameters of the 31P spin pair in Na4P2O7.10H2O representing our model case.     

Two-dimensional chemical shift/heteronuclear dipolar coupling spectra obtained with polarization inversion spin exchange at the magic angle and magic-angle sample spinning (PISEMAMAS)

High-resolution two-dimensional ¹⁵N chemical shift/¹H-¹⁵N dipolar coupling polarization inversion spin exchange at the magic angle (PISEMA) spectra of a polycrystalline sample of ¹⁵N-acetylvaline were obtained with and without magic-angle sample spinning. These spectra demonstrate the advantages of the PISEMA experiment over conventional approaches to separated local-field spectroscopy, especially the high resolution in the dipolar dimension where the spinning sidebands have uniformly narrow linewidths.     

H MAS and H[Na] double resonance NMR studies on the modification of surface hydroxyl groups of γ-alumina by sodium

The modification of surface hydroxyl groups with sodium in a series of Na₂CO₃-γ-Al₂O₃ catalysts was investigated as a function of both the Na₂CO₃ loading and the calcination temperature by means of ¹H magic angle spinning (MAS) and ¹H[²³Na] spin-echo double resonance NMR techniques. The ¹H NMR experiments revealed that sodium ions are homogeneously distributed over the alumina surface and closely coordinated with the surface hydroxyl groups. In the catalysts calcined at 250 °C, the acidic hydroxyl groups (with a chemical shift of 2.0 ppm) are preferentially associated with sodium ions at low Na₂CO₃ coverages (5 and 10%), while both the acidic and the basic (0 ppm) hydroxyl groups are accessible for sodium ions at high coverages (15 and 20%). The coordination causes a low-field shift of about 2 ppm in the ¹H MAS spectra, and a broad signal at 4.5 ppm appears. It is interesting that the 4.5 ppm signal is completely suppressed in the ¹H[²³Na] MAS experiments, providing direct evidence that a strong interaction exists between adsorbed sodium ions and the surface hydroxyl groups. Increasing the calcination temperature to 450 °C results in preferential removal of the acidic hydroxyl groups, and only the most basic hydroxyl groups remain when the calcination temperature is raised to 600 °C. This is attributed to the formation of the coordinated species which enhances the acidity of the surface hydroxyl groups and prompts their dehydroxylation, especially at high calcination temperature. Correlation of the ¹H MAS NMR results and catalytic activity measurements indicates that the basic hydroxyl groups are essential for the carbonyl sulfide hydrolysis reaction.     

非平衡磁控溅射双势阱静电波动及其共振耦合

非平衡磁控靶表面电场和磁场相互正交构成磁阱结构,磁控靶和与之平行的偏压基片之间形成了另一种势阱结构,等离子体静电波动在这两种势阱结构中形成耦合共振.采用Langmuir探针研究等离子体中参数和浮置电位信号的功率谱密度.典型放电条件下两种势阱结构中的本征频率分别约为30—50 kHz和10—20 kHz,两种势阱条件下根据声驻波共振模式计算的电子温度数值与实验结果相符合.