Chemical Shift Evolution Research Articles

Chemical bond reorganization in intramolecular proton transfer revealed by ultrafast X-ray photoelectron spectroscopy

Time-resolved X-ray photoelectron spectroscopy (TR-XPS) is used in a simulation study to monitor the excited state intramolecular proton transfer between oxygen and nitrogen atoms in 2-(iminomethyl)phenol. Real-time monitoring of the chemical bond breaking and forming processes is obtained through the time evolution of excited-state chemical shifts. By employing individual atomic probes of the proton donor and acceptor atoms, we predict distinct signals with opposite chemical shifts of the donor and acceptor groups during proton transfer. Details of the ultrafast bond breaking and forming dynamics are revealed by extending the classical electron spectroscopy chemical analysis to real time. Through a comparison with simulated time-resolved photoelectron spectroscopy at the valence level, the distinct advantage of TR-XPS is demonstrated thanks to its atom specificity.

SHARPER-DOSY: Sensitivity enhanced diffusion-ordered NMR spectroscopy

Since its discovery in mid-20th century, the sensitivity of Nuclear Magnetic Resonance (NMR) has increased steadily, in part due to the design of new, sophisticated NMR experiments. Here we report on a liquid-state NMR methodology that significantly increases the sensitivity of diffusion coefficient measurements of pure compounds, allowing to estimate their sizes using a much reduced amount of material. In this method, the diffusion coefficients are being measured by analysing narrow and intense singlets, which are invariant to magnetic field inhomogeneities. The singlets are obtained through signal acquisition embedded in short (<0.5 ms) spin-echo intervals separated by non-selective 180° or 90° pulses, suppressing the chemical shift evolution of resonances and their splitting due to J couplings. The achieved 10−100 sensitivity enhancement results in a 100−10000-fold time saving. Using high field cryoprobe NMR spectrometers, this makes it possible to measure a diffusion coefficient of a medium-size organic molecule in a matter of minutes with as little as a few hundred nanograms of material.

Solid‐state heteronuclear multiple‐quantum spectroscopy under a magic‐angle spinning frequency of 150kHz

AbstractWe hereby demonstrate that1H detected15N‐1H heteronuclear multiple‐quantum spectroscopy can be carried out at a magic angle spinning frequency of 150 kHz. While the15N‐1H multiple‐quantum coherences can be directly excited from the dipolar order created by the method of adiabatic demagnetization in the rotating frame, it is technically more advantageous to acquire the chemical shift evolution of the heteronuclear multiple‐quantum coherence by two separate chemical shift evolution periods for1H and15N. We also show that the heteronuclear multiple‐quantum correlation spectrum can be obtained by shearing the corresponding heteronuclear single‐quantum correlation spectrum.

Rapid Structural Analysis of Minute Quantities of Organic Solids by Exhausting 1H Polarization in Solid-State NMR Spectroscopy Under Fast Magic Angle Spinning.

Solid-state nuclear magnetic resonance (NMR) often suffers from significant limitations due to the inherent low signal sensitivity when low-γ nuclei are involved. Herein, we report an elegant solid-state NMR approach for rapid structural analysis of minute amounts of organic solids. By encoding staggered chemical shift evolution in the indirect dimension and staggered acquisition in the 1H dimension, a proton-detected homonuclear 1H/1H and heteronuclear 13C/1H chemical shift correlation (HETCOR) spectrum can be obtained simultaneously in a single experiment at a fast magic-angle-spinning (MAS) condition with barely increasing the experimental time. We further show that during the conventional 1H-detected HETCOR experimental time, multiple homonuclear 1H/1H correlation spectra can be recorded in addition to the HETCOR spectrum, enabling the determination of 1H-1H distances. We establish that abundant 1H polarization can be efficiently manipulated and fully utilized in proton-detected solid-state NMR spectroscopy for extraction of more critical structural information and thus reduction of the total experimental time.

Improving efficiency of measuring individual 1H coupling networks by pure shift 2D J-resolved NMR spectroscopy.

The 1H coupling networks, including 1H-1H correlation and J coupling values, provide the important information for structure elucidation and conformation analysis. However, the presence of a large number of couplings and the phase-twist lineshapes often prevents revealing 1H coupling networks. Here, we provide a clean absorption-mode 2D NMR method, SIMAJ (SImple Methods for 2D Absorption mode J-resolved spectrum), for a straightforward assignment and measurement of the coupling network involving the chosen proton. Relying on the pure shift element, 1H-1H couplings and chemical shift evolution are totally separately demonstrating along the F1 and F2 dimensions, respectively. Processing with a single experiment dataset and free of 45° spectral shearing, an absorption-mode 2D J-resolved spectrum can be reconstructed. Two pulse sequences were proposed as examples. The SIMAJ signal processing method will be a general procedure for obtaining absorption-mode lineshapes when analyzing the experiment datasets with chemical shifts and J coupling multiplets in the orthogonal dimensions. With excellent sensitivity, high spectral purity, and ability of easily identifying 1H-1H correlations, significant improvements are beneficial for structural, conformational, or complex composition analyses.

Pesticide use is linked to increased body size in a large mammalian carnivore

AbstractPollution and pesticide use have been linked to evolution of chemical resistance and phenotypic shifts in invertebrates, but less so in vertebrates. Here we provide evidence that poisoning directed towards a mammalian carnivore, the dingo (Canis dingo), is linked to an increase in dingo body mass. We compared the skull length of dingoes, a proxy for size, from three regions where dingo populations were controlled by distributing poisoned meat baits and an unbaited region, before and after the introduction of the toxin sodium fluoroacetate (Compound 1080). Following 1080 introduction, dingo skull length increased in baited regions but not in the unbaited region. We estimate that after 1080 introduction, the skull length of female and male dingoes in baited regions increased by 4.49 and 3.6 mm, respectively. This equates to a 1.02- and 0.86-kg increase in mean body masses of female and male dingoes, respectively. We hypothesize that dingo body size has increased in baited regions due to 1080 selecting for animals with larger body size or because a reduction in dingo abundance in baited areas may have removed constraints on growth imposed by intraspecific competition and prey availability. Our study provides evidence that pesticide use can prompt phenotypic change in comparatively large and long-lived large vertebrates.

Mirror symmetric broadband homodecoupled perfect echo spectroscopy

A new experiment for recording phase sensitive ω1-broadband homodecoupled TOCSY spectra is presented. The method is an extension of the already existing perfect echo (PE) filter, proposed to sample t1 chemical shift under sustained homodecoupling. The modification is made by attaching a time reversed perfect echo filter to a regular perfect echo scheme. Thus it becomes possible to acquire for longer t1 acquisition times without compromising the quality of homodecoupling. The mirror symmetric double perfect echo is implemented into the evolution period of a TOCSY experiment. A spin lock pulse purges undesired dispersive antiphase components at the end of the central t1 evolution period. Pure absorptive lineshapes with reduced proton spin multiplicities are obtained. The approach can be used in conjunction with real or constant time chemical shift evolution. In case of compounds with reduced T2 relaxation time, the real time approach is advisable, where the echo delays are an extension of the t1 evolution period. In this way, an unnecessary loss due to T2 relaxation is avoided. Using the pulse sequence in constant time mode at high t1max values gives ω1-homodecoupled TOCSY spectra without a significant dependence of the transfer amplitude on J. All experiments were carried out using non uniform sampling to decrease the measurement time. Experimental setup, advantages and limitations are discussed.

Cross-Peaks in Simple Two-Dimensional NMR Experiments from Chemical Exchange of Transverse Magnetisation.

Two‐dimensional correlation measurements such as COSY, NOESY, HMQC, and HSQC experiments are central to small‐molecule and biomolecular NMR spectroscopy, and commonly form the basis of more complex experiments designed to study chemical exchange occurring during additional mixing periods. However, exchange occurring during chemical shift evolution periods can also influence the appearance of such spectra. While this is often exploited through one‐dimensional lineshape analysis (“dynamic NMR”), the analysis of exchange across multiple chemical shift evolution periods has received less attention. Here we report that chemical exchange‐induced cross‐peaks can arise in even the simplest two‐dimensional NMR experiments. These cross‐peaks can have highly distorted phases that contain rich information about the underlying exchange process. The quantitative analysis of such peaks, from a single 2D spectrum, can provide a highly accurate characterisation of underlying exchange processes.

Cross‐Peaks in Simple Two‐Dimensional NMR Experiments from Chemical Exchange of Transverse Magnetisation

AbstractTwo‐dimensional correlation measurements such as COSY, NOESY, HMQC, and HSQC experiments are central to small‐molecule and biomolecular NMR spectroscopy, and commonly form the basis of more complex experiments designed to study chemical exchange occurring during additional mixing periods. However, exchange occurring during chemical shift evolution periods can also influence the appearance of such spectra. While this is often exploited through one‐dimensional lineshape analysis (“dynamic NMR”), the analysis of exchange across multiple chemical shift evolution periods has received less attention. Here we report that chemical exchange‐induced cross‐peaks can arise in even the simplest two‐dimensional NMR experiments. These cross‐peaks can have highly distorted phases that contain rich information about the underlying exchange process. The quantitative analysis of such peaks, from a single 2D spectrum, can provide a highly accurate characterisation of underlying exchange processes.

Multiplicity-edited 1 H-1 H TOCSY experiment.

A 1 H-1 H total correlation spectroscopy (TOCSY) experiment incorporating 13 C multiplicity information is proposed. In addition, broadband 1 H homodecoupling in the indirect dimension can be implemented using a perfect BIRD module that affords exclusive 1 H chemical shift evolution with full decoupling of all heteronuclear and homonuclear (including 2 JHH ) coupling constants. As a complement to the normal TOCSY and the recent PSYCHE-TOCSY experiments, this novel multiplicity-edited TOCSY experiment distinguishes between CH/CH3 (phased up) and CH2 (phased down) cross-peaks, which facilitates resonance analysis and assignment.

Formula omitted] gradient coherence selection using a tapered stripline

Pulsed-field gradients are common in modern liquid state NMR pulse sequences. They are often used instead of phase cycles for the selection of coherence pathways, thereby decreasing the time required for the NMR experiment. Soft off-resonance pulses with a B1 gradient result in a spatial encoding similar to that created by pulsed-field (B0) gradients. In this manuscript we show that pulse sequences with pulsed-field gradients can easily be converted to one which uses off-resonance B1 field gradient (OFFBEAT) pulses. The advantage of B1 gradient pulses for coherence selection is that the chemical shift evolution during the pulses is (partially) suppressed. Therefore no refocusing echos are required to correct for evolution during the gradient pulses. A tapered stripline is shown to be a convenient tool for creating a well-defined gradient in the B1 field strength. B1 gradient coherence selection using a tapered stripline is a simple and cheap alternative to B0 pulsed-field gradients.

A 13C-detected 15N double-quantum NMR experiment to probe arginine side-chain guanidinium 15N\u03b7 chemical shifts

Arginine side-chains are often key for enzyme catalysis, protein–ligand and protein–protein interactions. The importance of arginine stems from the ability of the terminal guanidinium group to form many key interactions, such as hydrogen bonds and salt bridges, as well as its perpetual positive charge. We present here an arginine 13Cζ-detected NMR experiment in which a double-quantum coherence involving the two 15Nη nuclei is evolved during the indirect chemical shift evolution period. As the precession frequency of the double-quantum coherence is insensitive to exchange of the two 15Nη; this new approach is shown to eliminate the previously deleterious line broadenings of 15Nη resonances caused by the partially restricted rotation about the Cζ–Nε bond. Consequently, sharp and well-resolved 15Nη resonances can be observed. The utility of the presented method is demonstrated on the L99A mutant of the 19 kDa protein T4 lysozyme, where the measurement of small chemical shift perturbations, such as one-bond deuterium isotope shifts, of the arginine amine 15Nη nuclei becomes possible using the double-quantum experiment.

Experimental technique for multi-qubit nuclear magnetic resonance system

With the development of quantum information and quantum computation science, quantum information processor has been widely used in different areas such as quantum simulation, quantum computation and quantum metrology and so on. To make quantum computer come true, we need to increase the number of controllable qubits of the system and improve the controllability to perform complex quantum manipulation. As a good experimental testbed for quantum information processing, nuclear magnetic resonance (NMR) spin system provides rich and sophisticated quantum control methods. In recent years a lot of multi-qubit experiments have been performed on the platform and a series of experimental technologies have been developed. In this paper, we firstly explain the difficulties of multi-qubit NMR experiments. Then by focusing on the experiment of 7-qubit labelled pseudo-pure state preparation and other relevant experiments, we review the technologies in multi-qubit experiments. Using the radio frequency selective method, the inhomogeneities of the radio frequency pulses are reduced and the spectral resolution is improved. After performing 1/2 spin selective sequence, we can regard the three methyl protons in the sample of crotonic acid as a single 1/2 spin nucleus and treat the whole molecule as a 7-qubit quantum information processor. We utilize Gauss pulses, Hermite pulses, composite pulses and gradient ascent pulse engineering (GRAPE) pulses to implement basic /2 and rotation operations. The GRAPE pulses are calculated by subspace GRAPE program to speed up the computation greatly. The errors of the basic pulses caused by chemical shift and J-coupling evolution can be estimated by the program of pulse compilation. It divides the errors of the pulses into a series of post-errors and pre-errors. A program of sequence compilation is used to eliminate the accumulated error of the whole pulse sequence, reduce the number of pulses and optimize the experimental duration. A variety of methods of quantum state tomography have been proposed to improve the efficiency of reading out information about quantum state. As an experimental example, we combine the above experimental technologies and perform the experiment of 7-qubit labelled pseudo-pure state preparation by using the method of cat state preparation. The sequence of cat state preparation consists of three steps:encoding procedure, phase cycling and decoding procedure. We use 14 experiments to realize the phase cycling and acquire the final 7-qubit labelled pseudo-pure state. The total duration of experimental sequence is about 132 ms. All the readout spectra have the similar shapes to the theoretical expectations. Finally we give an outlook for further research in this direction.

Constant-time 2D and 3D through-bond correlation NMR spectroscopy of solids under 60 kHz MAS.

Establishing connectivity and proximity of nuclei is an important step in elucidating the structure and dynamics of molecules in solids using magic angle spinning (MAS) NMR spectroscopy. Although recent studies have successfully demonstrated the feasibility of proton-detected multidimensional solid-state NMR experiments under ultrafast-MAS frequencies and obtaining high-resolution spectral lines of protons, assignment of proton resonances is a major challenge. In this study, we first re-visit and demonstrate the feasibility of 2D constant-time uniform-sign cross-peak correlation (CTUC-COSY) NMR experiment on rigid solids under ultrafast-MAS conditions, where the sensitivity of the experiment is enhanced by the reduced spin-spin relaxation rate and the use of low radio-frequency power for heteronuclear decoupling during the evolution intervals of the pulse sequence. In addition, we experimentally demonstrate the performance of a proton-detected pulse sequence to obtain a 3D (1)H/(13)C/(1)H chemical shift correlation spectrum by incorporating an additional cross-polarization period in the CTUC-COSY pulse sequence to enable proton chemical shift evolution and proton detection in the incrementable t1 and t3 periods, respectively. In addition to through-space and through-bond (13)C/(1)H and (13)C/(13)C chemical shift correlations, the 3D (1)H/(13)C/(1)H experiment also provides a COSY-type (1)H/(1)H chemical shift correlation spectrum, where only the chemical shifts of those protons, which are bonded to two neighboring carbons, are correlated. By extracting 2D F1/F3 slices ((1)H/(1)H chemical shift correlation spectrum) at different (13)C chemical shift frequencies from the 3D (1)H/(13)C/(1)H spectrum, resonances of proton atoms located close to a specific carbon atom can be identified. Overall, the through-bond and through-space homonuclear/heteronuclear proximities determined from the 3D (1)H/(13)C/(1)H experiment would be useful to study the structure and dynamics of a variety of chemical and biological solids.

Frequency-selective REDOR and spin-diffusion relays in uniformly labeled whole cells

Solid-state NMR is a powerful and non-perturbative method to measure and define chemical composition and architecture in bacterial cell walls, even in the context of whole cells. Most NMR studies on whole cells have used selectively labeled samples. Here, we introduce an NMR sequence relay using frequency-selective REDOR (fsREDOR) and spin diffusion elements to probe a unique amine contribution in uniformly 13C- and 15N-labeled Staphylococcus aureus whole cells that we attribute to the d-alanine of teichoic acid. In addition to the primary peptidoglycan structural scaffold, cell walls can contain significant amounts of teichoic acid that contribute to cell-wall function. When incorporated into teichoic acid, d-alanine is present as an ester, connected via its carbonyl to a ribitol carbon, and thus has a free amine. Teichoic acid d-Ala is removed during cell-wall isolations and can only be detected in the context of whole cells. The sequence presented here begins with fsREDOR and a chemical shift evolution period for 2D data acquisition, followed by DARR spin diffusion and then an additional fsREDOR period. fsREDOR elements were used for 13C observation to avoid complications from 13C–13C couplings due to uniform labeling and for 15N dephasing to achieve selectivity in the nitrogens serving as dephasers. The results show that the selected amine nitrogen of interest is near to teichoic acid ribitol carbons and also the methyl group carbon associated with alanine. In addition, its carbonyl is not significantly dephased by amide nitrogens, consistent with the expected microenvironment around teichoic acid.

High resolution 4D HPCH experiment for sequential assignment of (13)C-labeled RNAs via phosphodiester backbone.

The three-dimensional structure determination of RNAs by NMR spectroscopy requires sequential resonance assignment, often hampered by assignment ambiguities and limited dispersion of 1H and 13C chemical shifts, especially of C4′/H4′. Here we present a novel through-bond 4D HPCH NMR experiment involving phosphate backbone where C4′–H4′ correlations are resolved along the 1H3′–31P spectral planes. The experiment provides high peak resolution and effectively removes ambiguities encountered during assignments. Enhanced peak dispersion is provided by the inclusion of additional 31P and 1H3′ dimensions and constant-time evolution of chemical shifts. High spectral resolution is obtained by using non-uniform sampling in three indirect dimensions. The experiment fully utilizes the isotopic 13C-labeling with evolution of C4′ carbons. Band selective 13C inversion pulses are used to achieve selectivity and prevent signal dephasing due to the C4′–C3′ and C4′–C5′ homonuclear couplings. Multiple quantum line narrowing is employed to minimize sensitivity loses. The 4D HPCH experiment is verified and successfully applied to a non-coding 34-nt RNA consisting typical structure elements and a 14-nt RNA hairpin capped by cUUCGg tetraloop.

X-ray excited Auger transitions of Pu compounds

X-ray excited Pu core–valence–valence and core–core–valence Auger line-shapes were used in combination with the Pu 4f photoelectron peaks to characterize differences in the oxidation state and local electronic structure for Pu compounds. The evolution of the Pu 4f core-level chemical shift as a function of sputtering depth profiling and hydrogen exposure at ambient temperature was quantified. The combination of the core–valence–valence Auger peak energies with the associated chemical shift of the Pu 4f photoelectron line defines the Auger parameter and results in a reliable method for definitively determining oxidation states independent of binding energy calibration. Results show that PuO2, Pu2O3, PuH2.7, and Pu have definitive Auger line-shapes. These data were used to produce a chemical state (Wagner) plot for select plutonium oxides. This Wagner plot allowed us to distinguish between the trivalent hydride and the trivalent oxide, which cannot be differentiated by the Pu 4f binding energy alone.

Easy and unambiguous sequential assignments of intrinsically disordered proteins by correlating the backbone 15N or 13C' chemical shifts of multiple contiguous residues in highly resolved 3D spectra.

Sequential resonance assignment strategies are typically based on matching one or two chemical shifts of adjacent residues. However, resonance overlap often leads to ambiguity in resonance assignments in particular for intrinsically disordered proteins. We investigated the potential of establishing connectivity through the three-bond couplings between sequentially adjoining backbone carbonyl carbon nuclei, combined with semi-constant time chemical shift evolution, for resonance assignments of small folded and larger unfolded proteins. Extended sequential connectivity strongly lifts chemical shift degeneracy of the backbone nuclei in disordered proteins. We show here that 3D (H)N(COCO)NH and (HN)CO(CO)NH experiments with relaxation-optimized multiple pulse mixing correlate up to seven adjacent backbone amide nitrogen or carbonyl carbon nuclei, respectively, and connections across proline residues are also obtained straightforwardly. Multiple, recurrent long-range correlations with ultra-high resolution allow backbone (1)H(N), (15)N(H), and (13)C' resonance assignments to be completed from a single pair of 3D experiments.

NMR profiling of biomolecules at natural abundance using 2D 1H–15N and 1H–13C multiplicity-separated (MS) HSQC spectra

2D NMR 1H–X (X=15N or 13C) HSQC spectra contain cross-peaks for all XHn moieties. Multiplicity-edited1H–13C HSQC pulse sequences generate opposite signs between peaks of CH2 and CH/CH3 at a cost of lower signal-to-noise due to the 13C T2 relaxation during an additional 1/1JCH period. Such CHn-editing experiments are useful in assignment of chemical shifts and have been successfully applied to small molecules and small proteins (e.g. ubiquitin) dissolved in deuterated solvents where, generally, peak overlap is minimal. By contrast, for larger biomolecules, peak overlap in 2D HSQC spectra is unavoidable and peaks with opposite phases cancel each other out in the edited spectra. However, there is an increasing need for using NMR to profile biomolecules at natural abundance dissolved in water (e.g., protein therapeutics) where NMR experiments beyond 2D are impractical. Therefore, the existing 2D multiplicity-edited HSQC methods must be improved to acquire data on nuclei other than 13C (i.e.15N), to resolve more peaks, to reduce T2 losses and to accommodate water suppression approaches. To meet these needs, a multiplicity-separated1H–X HSQC (MS-HSQC) experiment was developed and tested on 500 and 700MHz NMR spectrometers equipped with room temperature probes using RNase A (14kDa) and retroviral capsid (26kDa) proteins dissolved in 95% H2O/5% D2O. In this pulse sequence, the 1/1JXH editing-period is incorporated into the semi-constant time (semi-CT) X resonance chemical shift evolution period, which increases sensitivity, and importantly, the sum and the difference of the interleaved 1JXH-active and the 1JXH-inactive HSQC experiments yield two separate spectra for XH2 and XH/XH3. Furthermore we demonstrate improved water suppression using triple xyz-gradients instead of the more widely used z-gradient only water-suppression approach.

Control of selective silicate glass coloration by gold metallic nanoparticles: structural investigation, growth mechanisms, and plasmon resonance modelization

Soda lime silicate oxide glasses are studied to perform coloration thanks to gold nanoparticles’ crystallization. This precipitation is conducted by chemical reduction of gold ions with stannous or antimony oxides as reducing agents. A control of the rendered coloration between blank to red shades has been obtained using Sb2O3 agent and appropriate thermal treatments. The glasses remain colorless while heating up to 450 °C. Structural glasses evolution is studied by MAS NMR spectroscopy of 29Si and 23Na nuclei to investigate the silicate network polymerization change and the modification of sodium/oxygen bond length versus nucleation state and growth of Au nanoparticles. A clear decrease of the Q2 species part is observed with nanoparticles growth confirmed by the evolution of chemical shift for 23Na resonance. A slight network polymerization is then showed independently of the only thermal treatment. This structural change could be induced by the antimony oxidation and change towards higher coordinations. Finally, the glasses chemical durability has been studied by leaching tests and shows lower alteration for colored glass. The optical spectroscopy applied to colored glasses has given rise to plasmon resonance phenomena at around 600 nm which is the typical surface plasmon resonance of gold for a refractive medium index of 1.5, with a shift of the resonance towards the higher wavelengths with increasing thermal treatment temperature. This shift is modelized by Drude and MIE approaches and confirms the trend observed by UV-visible measurement with an increasing absorption at the SPR correlated to a typical Ostwald growth mechanism according to the increase of the annealing temperature.