Rapid Chemical Exchange Research Articles

Controlled Sublimation Rate of Guest Drug from Polymorphic Forms of a Cyclodextrin-Based Polypseudorotaxane Complex and Its Correlation with Molecular Dynamics as Probed by Solid-State NMR.

Encapsulation of active pharmaceutical ingredients (APIs) in confined spaces has been extensively explored as it dramatically alters the molecular dynamics and physical properties of the API. Herein, we explored the effect of encapsulation on the molecular dynamics and physical stability of a guest drug, salicylic acid (SA), confined in the intermolecular spaces of γ-cyclodextrin (γ-CD) and poly(ethylene glycol) (PEG)-based polypseudorotaxane (PPRX) structure. The sublimation tendency of SA encapsulated in three polymorphic forms of the γ-CD/PEG-based PPRX complex, monoclinic columnar (MC), hexagonal columnar (HC), and tetragonal columnar (TC), was investigated. The SA sublimation rate was decreased by 3.0-6.6-fold and varied in the order of MC form > HC form > TC form complex. The 13C and 1H magic-angle spinning (MAS) solid-state nuclear magnetic resonance (NMR) spectra and 13C spin-lattice relaxation time (T1) indicated that the encapsulated SA molecules existed as the monomeric form, and its molecular mobility increased in the order of MC form > HC form > TC form complex. In the complexes, a rapid chemical exchange between two dynamic states of SA (free and bound) was suggested, with varying adsorption/desorption rates accounting for its distinct molecular mobility. This adsorption/desorption process was influenced by proton exchange at the interaction site and interaction strength of SA in the complexes, as evidenced by 1H MAS spectra and temperature dependency of the 13C carbonyl chemical shift. A positive correlation between the molecular mobility of SA and its sublimation rate was established. Moreover, the molecular mobility of γ-CD and PEG in the complexes coincided with that of SA, which can be explained by fast guest-driven dynamics. This is the first report on the stability improvement of an API through complexation in polymorphic supramolecular host structures. The relationship between the molecular dynamics and physical properties of encapsulated API will aid in the rational design of drug delivery systems.

Water Dynamics in Dextran-Based Hydrogel Micro/Nanoparticles Studied by NMR Diffusometry and Relaxometry.

Water dynamics in mesoporous dextran hydrogel micro/nanoparticles was investigated by means of nuclear magnetic resonance (NMR) techniques. High-resolution 1H NMR spectra and pulsed field gradient (PFG) NMR diffusometry measurements obtained on swollen state dextran micro/nanogel revealed the existence of different fractions of water molecules based on their interaction with the gel matrix. In addition to the translational diffusion of bulk water, two more diffusion processes characterized with self-diffusion coefficients 1 and 2 orders of magnitude smaller than that of bulk water were identified. 1H spin-lattice relaxation dispersion profiles obtained for a broad range of Larmor frequencies using fast field cycling (FFC) and conventional NMR relaxometry techniques allowed us to further clarify the mechanisms of molecular motion. According to the water proton pool fractions and associated self-diffusion coefficients, it is shown that the relaxation contribution associated with reorientation-mediated translational motions (RMTDs) dominates the relaxation dispersion observed at intermediate frequencies. At very low frequencies, the spin-lattice relaxation rate is dominated by the slow solid-gel dynamics probed by the water molecules interacting with the pores' surface hydroxyl groups due to the rapid chemical exchange between surface hydroxyl groups and free water. The correlation time for the thumbling-like motion of the dextran gel was found to be in the submillisecond range. The values of the self-diffusion and coherence lengths associated with motion of water molecules interacting with the solid-gel particles are consistent with the particle size and pore size distributions obtained for the studied dextran gels.

Accelerated and quantitative three-dimensional molecular MRI using a generative adversarial network.

To substantially shorten the acquisition time required for quantitative three-dimensional (3D) chemical exchange saturation transfer (CEST) and semisolid magnetization transfer (MT) imaging and allow for rapid chemical exchange parameter map reconstruction. Three-dimensional CEST and MT magnetic resonance fingerprinting (MRF) datasets of L-arginine phantoms, whole-brains, and calf muscles from healthy volunteers, cancer patients, and cardiac patients were acquired using 3T clinical scanners at three different sites, using three different scanner models and coils. A saturation transfer-oriented generative adversarial network (GAN-ST) supervised framework was then designed and trained to learn the mapping from a reduced input data space to the quantitative exchange parameter space, while preserving perceptual and quantitative content. The GAN-ST 3D acquisition time was 42-52 s, 70% shorter than CEST-MRF. The quantitative reconstruction of the entire brain took 0.8 s. An excellent agreement was observed between the ground truth and GAN-based L-arginine concentration and pH values (Pearson's r > 0.95, ICC > 0.88, NRMSE < 3%). GAN-ST images from a brain-tumor subject yielded a semi-solid volume fraction and exchange rate NRMSE of and , respectively, and SSIM of and , respectively. The mapping of the calf-muscle exchange parameters in a cardiac patient, yielded NRMSE < 7% and SSIM > 94% for the semi-solid exchange parameters. In regions with large susceptibility artifacts, GAN-ST has demonstrated improved performance and reduced noise compared to MRF. GAN-ST can substantially reduce the acquisition time for quantitative semi-solid MT/CEST mapping, while retaining performance even when facing pathologies and scanner models that were not available during training.

Polarizing insensitive nuclei at ultralow magnetic fields using parahydrogen: A facile route to optimize adiabatic magnetic field sweeps.

Parahydrogen induced polarization (PHIP) provides a powerful tool to enhance inherently weak nuclear magnetic resonance signals, particularly in biologically relevant compounds. The initial source of PHIP is the non-equilibrium spin order of parahydrogen, i.e., dihydrogen, where the two protons make up a singlet spin state. Conversion of this spin order into net magnetization of magnetic heteronuclei, e.g., 13C, provides one of the most efficient ways to exploit PHIP. We propose a facile route to increase the performance of PHIP transfer in experiments with adiabatic sweeps of the ultralow magnetic field. To date, this technique yields the highest efficiency of PHIP transfer, yet, it has been mostly utilized with linear field sweeps, which does not consider the underlying spin dynamics, resulting in sub-optimal polarization. This issue was previously addressed by using the "constant" adiabaticity method, which, however, requires extensive calculations for large spin systems. In this work, the field sweep is optimized by utilizing the field dependence of the average 13C polarization. Both the experimental detection and the numerical simulation of this dependence are straightforward, even for complex multi-spin systems. This work provides a comprehensive survey of PHIP transfer dynamics at ultralow fields for two molecular systems that are relevant for PHIP, namely, maleic acid and allyl pyruvate. The proposed optimization allowed us to increase the resulting 13C polarization in 13C-allyl pyruvate from 6.8% with a linear profile to 8.7% with an "optimal" profile. Such facile optimization routines are valuable for adiabatic experiments in complex spin systems undergoing rapid relaxation or chemical exchange.

An end-to-end AI-based framework for automated discovery of rapid CEST/MT MRI acquisition protocols and molecular parameter quantification (AutoCEST).

PurposeTo develop an automated machine‐learning‐based method for the discovery of rapid and quantitative chemical exchange saturation transfer (CEST) MR fingerprinting acquisition and reconstruction protocols.MethodsAn MR physics‐governed AI system was trained to generate optimized acquisition schedules and the corresponding quantitative reconstruction neural network. The system (termed AutoCEST) is composed of a CEST saturation block, a spin dynamics module, and a deep reconstruction network, all differentiable and jointly connected. The method was validated using a variety of chemical exchange phantoms and in vivo mouse brains at 9.4T.ResultsThe acquisition times for AutoCEST optimized schedules ranged from 35 to 71 s, with a quantitative image reconstruction time of only 29 ms. The resulting exchangeable proton concentration maps for the phantoms were in good agreement with the known solute concentrations for AutoCEST sequences (mean absolute error = 2.42 mM; Pearson’s r=0.992, p<0.0001), but not for an unoptimized sequence (mean absolute error = 65.19 mM; Pearson’s r=‐0.161, p=0.522). Similarly, improved exchange rate agreement was observed between AutoCEST and quantification of exchange using saturation power (QUESP) methods (mean absolute error: 35.8 Hz, Pearson’s r=0.971, p<0.0001) compared to an unoptimized schedule and QUESP (mean absolute error = 58.2 Hz; Pearson’s r=0.959, p<0.0001). The AutoCEST in vivo mouse brain semi‐solid proton volume fractions were lower in the cortex (12.77% ± 0.75%) compared to the white matter (19.80% ± 0.50%), as expected.ConclusionAutoCEST can automatically generate optimized CEST/MT acquisition protocols that can be rapidly reconstructed into quantitative exchange parameter maps.

NHC‐Stabilized Parent Phosphinidene Adducts of Metal(II) Hexamethyldisilazanides of Manganese – Cobalt and Their Lability in Solution

AbstractThe behavior of the N‐heterocyclic carbene (NHC) stabilized ‘parent’ phosphinidene [(sIDipp)PH] (Dipp=2,6‐di‐isopropylphenyl) towards 3d‐metal(II) hexamethyldisilazanides is described. This leads to formation of the three‐coordinated phosphinidene adducts [M{PH(sIDipp)}{N(SiMe3)2}2] (M=Mn−Co), as evidenced by X‐Ray diffraction analysis. In solution, all compounds exhibit strong paramagnetism that limited NMR spectroscopic examinations to cobalt. Here it shows that the system is coordinatively highly fluxional, which is mitigtated upon cooling. At low temperatures, small amounts of free phosphinidene becomes further evident by 31P NMR spectroscopy that implicates in part ligand coordination competes with [M{N(SiMe3)2}2] dimerisation and an overall weak binding of the phosphinidene ligand. The rapid chemical exchange is evidenced in the presence of an excess of phosphinidene or a phosphine (PCy3), whereas diethyl ether proved as a stronger donor ligand.

Using bidirectional chemical exchange for improved hyperpolarized [13 C]bicarbonate pH imaging.

Rapid chemical exchange can affect SNR and pH measurement accuracy for hyperpolarized pH imaging with [13 C]bicarbonate. The purpose of this work was to investigate chemical exchange effects on hyperpolarized imaging sequences to identify optimal sequence parameters for high SNR and pH accuracy. Simulations were performed under varying rates of bicarbonate-CO2 chemical exchange to analyze exchange effects on pH quantification accuracy and SNR under different sampling schemes. Four pulse sequences, including 1 new technique, a multiple-excitation 2D EPI (multi-EPI) sequence, were compared in phantoms using hyperpolarized [13 C]bicarbonate, varying parameters such as tip angles, repetition time, order of metabolite excitation, and refocusing pulse design. In vivo hyperpolarized bicarbonate-CO2 exchange measurements were made in transgenic murine prostate tumors to select in vivo imaging parameters. Modeling of bicarbonate-CO2 exchange identified a multiple-excitation scheme for increasing CO2 SNR by up to a factor of 2.7. When implemented in phantom imaging experiments, these sampling schemes were confirmed to yield high pH accuracy and SNR gains. Based on measured bicarbonate-CO2 exchange in vivo, a 47% CO2 SNR gain is predicted. The novel multi-EPI pulse sequence can boost CO2 imaging signal in hyperpolarized 13 C bicarbonate imaging while introducing minimal pH bias, helping to surmount a major hurdle in hyperpolarized pH imaging.

Hydrogen dynamics features in BaZr1 − x Sc x O3 − x/2(OH) y : high-temperature 1H NMR studies

The 1H nuclear magnetic resonance measurements were carried out to study proton dynamics features in the hydrated scandium-doped barium zirconate, BaZr1 − xScxO3 − x/2(OH)y, with x = 0.2 and 0.4 in the temperature range 300–600 K. The obtained data evidence a fast proton motion in both samples, which is characterized by the jump frequency of about 108 s−1 at 450 K. However, the microscopic nature of hydrogen diffusion in these two samples is quite different. For the sample with x = 0.2, hydrogen motion mechanisms are defined by a rapid chemical exchange between Sc–OH–Zr and Zr–OH–Zr positions. The estimated value of activation energy for hydrogen diffusion of about 0.5 eV is determined by the energy barrier produced by Sc3+ ion. The increase of Sc concentration to x = 0.4 leads to the drastic changes of sample properties. Experimental results allow to assume the formation of nanoscaled Sc-rich domains and the decomposition of hydrogen sublattice with formation of two proton subsystems: the protons in Zr–OH–Zr coordinations and those concentrated in Sc–OH–Sc environments. The proton motion in both these subsystems is rather fast, but the chemical exchange between them is highly suppressed. Hydrogen motion inside Sc-rich environments has most likely localized nature. Our estimates yield the energy values of about 0.25 and 0.55 eV for hydrogen motion in “free lattice” and in Sc-rich clusters, respectively. The Sc-rich domains can retain hydrogen up to 600 K.

Computational Infrared and Two-Dimensional Infrared Photon Echo Spectroscopy of Both Wild-Type and Double Mutant Myoglobin-CO Proteins

The CO stretching mode of both wild-type and double mutant ( T67R / S92D ) MbCO (carbonmonoxymyoglobin) proteins is an ideal infrared (IR) probe for studying the local electrostatic environment inside the myoglobin heme pocket. Recently, to elucidate the conformational switching dynamics between two distinguishable states, extensive IR absorption, IR pump-probe, and two-dimensional (2D) IR spectroscopic studies for various mutant MbCO's have been performed by the Fayer group. They showed that the 2D IR spectroscopy of the double mutant, which has a peroxidase enzyme activity, reveals a rapid chemical exchange between two distinct states, whereas that of the wild-type does not. Despite the fact that a few simulation studies on these systems were already performed and reported, such complicated experimental results have not been fully reproduced nor described in terms of conformational state-to-state transition processes. Here, we first develop a distributed vibrational solvatochromic charge model for describing the CO stretch frequency shift reflecting local electric potential changes. Then, by carrying out molecular dynamic simulations of the two MbCO's and examining their CO frequency trajectories, it becomes possible to identify a proper reaction coordinate consisting of His64 imidazole ring rotation and its distance to the CO ligand. From the 2D surfaces of the resulting potential of mean forces, the spectroscopically distinguished A1 and A3 states of the wild-type as well as two more substates of the double mutant are identified and their vibrational frequencies and distributions are separately examined. Our simulated IR absorption and 2D IR spectra of the two MbCO's are directly compared with the previous experimental results reported by the Fayer group. The chemical exchange rate constants extracted from the two-state kinetic analyses of the simulated 2D IR spectra are in excellent agreement with the experimental values. On the basis of the quantitative agreement between the simulated spectra and experimental ones, we further examine the conformational differences in the heme pockets of the two proteins and show that the double mutation, T67R / S92D , suppresses the A1 population, restricts the imidazole ring rotation, and increases hydrogen-bond strength between the imidazole Nε-H and the oxygen atom of the CO ligand. It is believed that such delicate change of distal His64 imidazole ring dynamics induced by the double mutation may be responsible for its enhanced peroxidase catalytic activity as compared to the wild-type myoglobin.

Contributions of chemical and diffusive exchange to T1ρ dispersion

Variations in local magnetic susceptibility may induce magnetic field gradients that affect the signals acquired for MR imaging. Under appropriate diffusion conditions, such fields produce effects similar to slow chemical exchange. These effects may also be found in combination with other chemical exchange processes at multiple time scales. We investigate these effects with simulations and measurements to determine their contributions to rotating frame (R1ρ ) relaxation in model systems. Simulations of diffusive and chemical exchange effects on R1ρ dispersion were performed using the Bloch equations. Additionally, R1ρ dispersion was measured in suspensions of Sephadex and latex beads with varying spin locking fields at 9.4 T. A novel analysis method was used to iteratively fit for apparent chemical and diffusive exchange rates with a model by Chopra et al. Single- and double-inflection points in R1ρ dispersion profiles were observed, respectively, in simulations of slow diffusive exchange alone and when combined with rapid chemical exchange. These simulations were consistent with measurements of R1ρ in latex bead suspensions and small-diameter Sephadex beads that showed single- and double-inflection points, respectively. These observations, along with measurements following changes in temperature and pH, are consistent with the combined effects of slow diffusion and rapid -OH exchange processes.

Characterization and dynamics of substituted ruthenacyclobutanes relevant to the olefin cross-metathesis reaction.

The reaction of the phosphonium alkylidene [(H(2)IMes)RuCl(2)=CHP(Cy)(3))](+) BF(4)(-) with propene, 1-butene, and 1-hexene at -45 °C affords various substituted, metathesis-active ruthenacycles. These metallacycles were found to equilibrate over extended reaction times in response to decreases in ethylene concentrations, which favored increased populations of α-monosubstituted and α,α'-disubstituted (both cis and trans) ruthenacycles. On an NMR time scale, rapid chemical exchange was found to preferentially occur between the β-hydrogens of the cis and trans stereoisomers prior to olefin exchange. Exchange on an NMR time scale was also observed between the α- and β-methylene groups of the monosubstituted ruthenacycle (H(2)IMes)Cl(2)Ru(CHRCH(2)CH(2)) (R = CH(3), CH(2)CH(3), (CH(2))(3)CH(3)). EXSY NMR experiments at -87 °C were used to determine the activation energies for both of these exchange processes. In addition, new methods have been developed for the direct preparation of metathesis-active ruthenacyclobutanes via the protonolysis of dichloro(1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene)(benzylidene) bis(pyridine)ruthenium(II) and its 3-bromopyridine analogue. Using either trifluoroacetic acid or silica-bound toluenesulfonic acid as the proton source, the ethylene-derived ruthenacyclobutane (H(2)IMes)Cl(2)Ru(CH(2)CH(2)CH(2)) was observed in up to 98% yield via NMR at -40 °C. On the basis of these studies, mechanisms accounting for the positional and stereochemical exchange within ruthenacyclobutanes are proposed, as well as the implications of these dynamics toward olefin metathesis catalyst and reaction design are described.

Mixed-ligand complexes of zinc(II), cobalt(II) and cadmium(II) with sulfur, nitrogen and oxygen ligands. Analysis of the solid state structure and solution behavior. Implications for metal ion substitution in alcohol dehydrogenase

In this paper we characterize new, mixed ligand complexes of zinc(II), cobalt(II) and cadmium(II) with tri- tert-butoxysilanethiolate and 2-(2′-hydroxyethyl)pyridine ligands. Due to the chelating versus non-chelating behavior of 2-(2′-hydroxyethyl)pyridine ligand we have obtained an interesting structural variety in the studied system. The presented coordination patterns together with the results of NMR studies have been used to illustrate a rapid chemical exchange undergoing in methanolic solutions of zinc(II) and cadmium(II) complexes. UV–Vis spectra of cobalt(II) species have also evidenced an exchange in the case of cobalt(II) complex. The relative strength of hydrogen bond formed by hydroxyl group bonded to Zn(II), Co(II) or Cd(II) was evaluated by analysis of structural parameters and position of the OH stretching vibrations in the FT-IR spectra of the complexes in solid state. The data were compared with the activity of zinc (native) alcohol dehydrogenase and alcohol dehydrogenase substituted with cobalt and cadmium ions. The enthalpies of proton abstraction in zinc and cobalt complexes were calculated and found to be very similar. The attempt to apply zinc tri- tert-butoxysilanethiolate as a catalyst in the biomimetic reaction of reduction of N-benzylnicotinamide chloride by ethanol was unsuccessful.

Effects of frozen storage and sample temperature on water compartmentation and multiexponential transverse relaxation in cartilage

Multiexponential transverse relaxation in tissue has been interpreted as a marker of water compartmentation. Articular cartilage has been reported to exhibit such relaxation in several studies, with the relative contributions of tissue heterogeneity and tissue microstructure remaining unspecified. In bovine nasal cartilage, conflicting data regarding the existence of multiexponential relaxation have been reported. Imaging and analysis artifacts as well as rapid chemical exchange between tissue compartments have been identified as potential causes for this discrepancy. Here, we find that disruption of cartilage microstructure by freeze-thawing can greatly alter the character of transverse relaxation in this tissue. We conclude that fresh cartilage exhibits multiexponential relaxation based upon its microstructural water compartments, but that multiexponentiality can be lost or rendered undetectable by freeze-thawing. In addition, we find that increasing chemical exchange by raising sample temperature from 4°C to 37°C does not substantially limit the ability to detect multiexponential relaxation.

CEST‐FISP: A novel technique for rapid chemical exchange saturation transfer MRI at 7 T

Chemical exchange saturation transfer (CEST) and magnetization transfer techniques provide unique and potentially quantitative contrast mechanisms in multiple MRI applications. However, the in vivo implementation of these techniques has been limited by the relatively slow MRI acquisition techniques, especially on high-field MRI scanners. A new, rapid CEST-fast imaging with steady-state free precession technique was developed to provide sensitive CEST contrast in ∼20 sec. In this study at 7 T with in vitro bovine glycogen samples and initial in vivo results in a rat liver, the CEST-fast imaging with steady-state free precession technique was shown to provide equivalent CEST sensitivity in comparison to a conventional CEST-spin echo acquisition with a 50-fold reduction in acquisition time. The sensitivity of the CEST-fast imaging with steady-state free precession technique was also shown to be dependent on k-space encoding with centric k-space encoding providing a 30-40% increase in CEST sensitivity relative to linear encoding for 256 or more k-space lines. Overall, the CEST-fast imaging with steady-state free precession acquisition technique provides a rapid and sensitive imaging platform with the potential to provide quantitative CEST and magnetization transfer imaging data.

HA-detected experiments for the backbone assignment of intrinsically disordered proteins

We propose a new alpha proton detection based approach for the sequential assignment of natively unfolded proteins. The proposed protocol superimposes on following features: HA-detection (1) enables assignment of natively unfolded proteins at any pH, i.e., it is not sensitive to rapid chemical exchange undergoing in natively unfolded proteins even at moderately high pH. (2) It allows straightforward assignment of proline-rich polypeptides without additional proline-customized experiments. (3) It offers more streamlined and less ambiguous assignment based on solely intraresidual (15)N(i)-(13)C'(i)-H(alpha)(i) (or (15)N(i)-(13)C(alpha)(i)-H(alpha)(i)) and sequential (15)N(i + 1)-(13)C'(i)-H(alpha)(i) (or (15)N(i + 1)-(13)C(alpha)(i)-H(alpha)(i)) correlation experiments together with efficient use of chemical shifts of (15)N and (13)C' nuclei, which show smaller dependence on residue type. We have tested the proposed protocol on two proteins, small globular 56-residue GB1, and highly disordered, proline-rich 47-residue fifth repeat of EspF(U). Using the proposed approach, we were able to assign 90% of (1)H(alpha), (13)C(alpha), (13)C', (15)N chemical shifts in EspF(U). We reckon that the HA-detection based strategy will be very useful in the assignment of natively unfolded proline-rich proteins or polypeptide chains.

Homoleptic Zinc(II) Complexes with First and Second Coordination Shells of 5-tert-Butylpyrazole

Reaction of hydrated Zn[NO3]2 or Zn[BF4]2 with four or more equivalents of 3{5}-tert-butylpyrazole (L(tBu)) yields [Zn(L(tBu))4]X2 (X- = NO3- or BF4-). The nitrate complex contains C2-symmetric four-coordinate zinc(II) centers with a slightly flattened tetrahedral geometry, and each nitrate anion hydrogen bonds to two pyrazole N-H groups. Similar reactions with Zn[ClO4]2 or ZnCl2 in the presence of 2 equiv of AgPF6 or AgSbF6 yield instead [{Zn(L(tBu))4}(L(tBu))4][ClO4]2 and [{Zn(L(tBu))4}(L(tBu))2]Y2 (Y- = PF6- or SbF6-). Crystals of [{Zn(L(tBu))4}(L(tBu))4][ClO4]2 are composed of discrete [{Zn(L(tBu))4}(L(tBu))4]2+ supramolecules that are formed from N-H...N hydrogen bonding between zinc-bound and uncoordinated pyrazole rings. The [{Zn(L(tBu))4}(L(tBu))4]2+ moieties are linked into planar 4(4) nets by hydrogen bonding to bridging ClO4- anions. The ClO4- ions are almost perfectly encapsulated in near-spherical cavities of approximate dimensions 5.0 x 5.0 x 4.5 A that are formed by two interlocked supramolecular dications. Similarly, [{Zn(L(tBu))4}(L(tBu))2][PF6]2 crystallizes as discrete supramolecules in the crystal with the PF6- anions occupying a shallow bowl-shaped cavity on the surface of the complex that is formed by two zinc-bound and one uncoordinated pyrazole ligands. (1)H NMR and IR studies of [{Zn(L(tBu))4}(L(tBu))4][ClO4]2 in CD2Cl2 imply that the second-sphere L(tBu) ligands dissociate from the [Zn(L(tBu))4]2+ center in this solvent and that free and metal-bound L(tBu) are in rapid chemical exchange.

Cation Binding in Na,K-ATPase, Investigated by 205Tl Solid-State NMR Spectroscopy

Cation binding to Na,K-ATPase is characterized in native membranes at room temperature by solid-state NMR spectroscopy using the K(+) congener (205)Tl. It has been demonstrated that the signals from occluded Tl(+) and nonspecifically bound Tl(+) can be detected and distinguished by NMR. Effects of dipole-dipole coupling between (1)H and (205)Tl in the occlusion sites show that the ions are rigidly bound, rather than just occluded. Furthermore, a low chemical shift suggests occlusion site geometries with a relatively small contribution from carboxylate and hydroxyl groups. Nonspecific binding of Tl(+) is characterized by rapid chemical exchange, in agreement with the observed low binding affinity.

Enhancement of SQUID-detected NMR signals with hyperpolarized liquid 129Xe in a 1 microT magnetic field.

We report an enhancement of proton NMR signals by a factor of 10(6) by cross polarization with hyperpolarized liquid 129Xe in an ultralow magnetic field of 1 microT. The NMR signals from cyclopentane, acetone, and methanol are detected using a commercial high-T(c) SQUID magnetometer with a signal-to-noise ratio of up to 1000 from a single 90 degrees tipping pulse. This technique allows a wide range of low-field NMR measurements and is promising for the detection of intermolecular scalar spin-spin couplings. Scalar intermolecular couplings can produce a shift of the average NMR frequency in a hyperpolarized sample even in the presence of rapid chemical exchange.

Hydration structure and dynamics in pullulan aqueous solution based on 1H NMR relaxation time

Dynamic properties of water and polymer chain proton in pullulan/H 2O systems in aqueous solutions and in frozen states were analyzed based on T 2 relaxation times in 1H-NMR and DSC. Two relaxing species with different T 2 detected in the CPMG pulse sequence were assigned to inert polymer protons with the shorter T 2 and to water protons with the longer T 2 by using deuterated pullulan solutions in D 2O. It has been proved that hydration water and free water undergoes rapid exchange in pullulan aqueous solutions. In the frozen state at −11°C, protons in ice crystals (T 2∼17 μs), protons in mobile water ( T 2>ms) and inert and labile protons in polymer chains (T 2∼0.1 ms) were distinguished in FID curves measured by using the solid echo pulse sequence. With increase in temperature, the inert protons and the labile protons show different mobility, and the inert protons are separately observable from the labile protons in the Carr–Percell–Meiboom–Gill (CPMG) method at temperatures higher than 0°C and the labile protons become mixed with water protons by rapid chemical exchange. These findings indicate the extremely high flexibility of the pullulan chain in aqueous solution.

Solid and Solution State NMR Spectra and the Structure of the Gallium Citrate Complex (NH4)3[Ga(C6H5O7)2]·4H2O

A gallium citrate complex (NH4)3[Ga(C6H5O7)2]·4H2O (1) has been isolated from an aqueous mixture of gallium nitrate and citric acid at pH = 9. In the solid state 1 is monomeric and includes the near centrosymmetric ion [Ga(C6H5O7)2]3− in the asymmetric unit, with each tridentate citrate coordinating to gallium through an alkoxide oxygen, an oxygen of the central carboxyl group and an oxygen of a terminal carboxyl group. The second terminal carboxyl group is not bound to the metal centre. The two alkoxide oxygen−gallium distances in the asymmetric unit are significantly shorter than the four carboxyl oxygen−gallium distances, indicating a stronger bond for the former. The number and intensities of the resolved resonances in the solid state 13C MAS/NMR spectrum are consistent with the X-ray structure. In aqueous solution near neutral pH (5.5 to 6.4) the complex partially dissociates to give an equilibrium of the 1:1 and 1:2 gallium/citrate species together with free citrate. Two sets of resonances are observed in both the 1H and 13C NMR spectra of solutions of the complex and these correspond to bound and free citrate; there is no resolution of resonances for the 1:1 and 1:2 species. The relative strength of the alkoxide oxygen−gallium bond in the metal-bound citrate ligand leads to a slow intermolecular chemical exchange situation (rate < ca. 12 s−1) between the bound and free ligand. The carboxylate oxygen−gallium bonds are quite labile and relatively rapid intramolecular chemical exchange occurs (rate > ca. 700 s−1) between the metal-bound and pendant terminal carboxyl groups, averaging their 1H and 13C NMR signals. The diffusion coefficients for the free and complexed citrate measured from the 1H NMR spectra do not appear to be complicated by chemical exchange effects and, as expected, the diffusion coefficient for the complexed ligand is smaller than for the free ligand. In solution in the intermediate pH range (4.8 to 6.4) a new 71Ga signal is observed at ca. δ = 27 (Δν1/2 ≈︁ 10.6 kHz) which is assigned to the gallium citrate complexed species.