Pulsed Electron Paramagnetic Resonance Measurements Research Articles

Pulse EPR spectroscopy and molecular modeling reveal the origins of the local heterogeneity of dietary fibers

Optimizing human diet by including dietary fibers would be more efficient when the fibers' chain interactions with other molecules are understood in depth. Thereby, it is important to develop methods for characterizing the fiber chain to be able to monitor its structural alterations upon intermolecular interactions. Here, we demonstrate the utility of the electron paramagnetic resonance (EPR) spectroscopy, complemented by simulations in probing the atomistic details of the chain conformations for spin-labeled fibers. Barley β-glucan, a native polysaccharide with linear chain, was utilized as a test fiber system to demonstrate the technique's capabilities. Pulse dipolar EPR data show good agreement with results of the fiber chain modeling, revealing sinuous chain conformations and providing polymer shape descriptors: the gyration tensor, spin-spin distance distribution function, and information about proton density near the spin probe. Results from EPR measurements point to the fiber aggregation in aqueous solution, which agrees with the results of the dynamic light scattering. We propose that the combination of pulse EPR measurements with modeling can be a perfect experimental tool for in-depth structural investigation of dietary fibers and their interaction under such conditions, and that the presented methodology can be extended to other weakly ordered or disordered macromolecules.

Generation and Characterization of a Tetraradical Embedded in a Curved Cyclic Paraphenylene Unit.

Unique spin-spin (magnetic) interactions, ring-size effects on ground-state spin multiplicity, and in-plane aromaticity has been found in localized 1,3-diradicals embedded in curved benzene structures such as cycloparaphenylene (CPP). In this study, we characterized the magnetic interactions in a tetraradical consisting of two localized 1,3-diradical units connected by p-quaterphenyl within a curved CPP skeleton by electron paramagnetic resonance (EPR) spectroscopy and quantum chemical calculations. Persistent triplet species with zero-field splitting parameters similar to those of a triplet 1,3-diphenylcyclopentane-1,3-diyl diradical were observed by continuous wave (CW) or pulsed X-band EPR measurements. The quintet state derived from the ferromagnetic interaction between the two triplet diradical moieties was not detected at 20 K under glassy matrix conditions. At the B3LYP/6-31G(d) level of theory, the singlet state was lower in energy than the triplet and quintet states. These findings will aid in the development of open-shell species for material science application.

In situ amplification of spin echoes within a kinetic inductance parametric amplifier.

The use of superconducting microresonators together with quantum-limited Josephson parametric amplifiers has enhanced the sensitivity of pulsed electron spin resonance (ESR) measurements by more than four orders of magnitude. So far, the microwave resonators and amplifiers have been designed as separate components due to the incompatibility of Josephson junction-based devices with magnetic fields. This has produced complex spectrometers and raised technical barriers toward adoption of the technique. Here, we circumvent this issue by coupling an ensemble of spins directly to a weakly nonlinear and magnetic field-resilient superconducting microwave resonator. We perform pulsed ESR measurements with a 1-pL mode volume containing 6 × 107 spins and amplify the resulting signals within the device. When considering only those spins that contribute to the detected signals, we find a sensitivity of [Formula: see text] for a Hahn echo sequence at a temperature of 400 mK. In situ amplification is demonstrated at fields up to 254 mT, highlighting the technique's potential for application under conventional ESR operating conditions.

NMR and EPR reveal a compaction of the RNA-binding protein FUS upon droplet formation.

Many RNA-binding proteins undergo liquid-liquid phase separation, which underlies the formation of membraneless organelles, such as stress granules and P-bodies. Studies of the molecular mechanism of phase separation in vitro are hampered by the coalescence and sedimentation of organelle-sized droplets interacting with glass surfaces. Here, we demonstrate that liquid droplets of fused in sarcoma (FUS)-a protein found in cytoplasmic aggregates of amyotrophic lateral sclerosis and frontotemporal dementia patients-can be stabilized in vitro using an agarose hydrogel that acts as a cytoskeleton mimic. This allows their spectroscopic characterization by liquid-phase NMR and electron paramagnetic resonance spectroscopy. Protein signals from both dispersed and condensed phases can be observed simultaneously, and their respective proportions can be quantified precisely. Furthermore, the agarose hydrogel acts as a cryoprotectant during shock-freezing, which facilitates pulsed electron paramagnetic resonance measurements at cryogenic temperatures. Surprisingly, double electron-electron resonance measurements revealed a compaction of FUS in the condensed phase.

Going the dHis-tance: Site-Directed Cu2+ Labeling of Proteins and Nucleic Acids.

In this Account, we showcase site-directed Cu2+ labeling in proteins and DNA, which has opened new avenues for the measurement of the structure and dynamics of biomolecules using electron paramagnetic resonance (EPR) spectroscopy. In proteins, the spin label is assembled in situ from natural amino acid residues and a metal complex and requires no post-expression synthetic modification or purification procedures. The labeling scheme exploits a double histidine (dHis) motif, which utilizes endogenous or site-specifically mutated histidine residues to coordinate a Cu2+ complex. Pulsed EPR measurements on such Cu2+-labeled proteins potentially yield distance distributions that are up to 5 times narrower than the common protein spin label-the approach, thus, overcomes the inherent limitation of the current technology, which relies on a spin label with a highly flexible side chain. This labeling scheme provides a straightforward method that elucidates biophysical information that is costly, complicated, or simply inaccessible by traditional EPR labels. Examples include the direct measurement of protein backbone dynamics at β-sheet sites, which are largely inaccessible through traditional spin labels, and rigid Cu2+-Cu2+ distance measurements that enable higher precision in the analysis of protein conformations, conformational changes, interactions with other biomolecules, and the relative orientations of two labeled protein subunits. Likewise, a Cu2+ label has been developed for use in DNA, which is small, is nucleotide independent, and is positioned within the DNA helix. The placement of the Cu2+ label directly reports on the biologically relevant backbone distance. Additionally, for both of these labeling techniques, we have developed models for interpretation of the EPR distance information, primarily utilizing molecular dynamics (MD) simulations. Initial results using force fields developed for both protein and DNA labels have agreed with experimental results, which has been a major bottleneck for traditional spin labels. Looking ahead, we anticipate new combinations of MD and EPR to further our understanding of protein and DNA conformational changes, as well as working synergistically to investigate protein-DNA interactions.

Deep Insights into Heavy Oil Upgrading Using Supercritical Water by a Comprehensive Analysis of GC, GC-MS, NMR, and SEM-EDX with the Aid of EPR as a Complementary Technical Analysis.

Upgrading of heavy oil in supercritical water (SCW) was analyzed by a comprehensive analysis of GC, GC–MS, NMR, and SEM–EDX with the aid of electron paramagnetic resonance (EPR) as a complementary technical analysis. The significant changes in the physical properties and chemical compositions reveal the effectiveness of heavy oil upgrading by SCW. Especially, changes of intensities of conventional EPR signals from free radicals (FRs) and paramagnetic vanadyl complexes (VO2+) with SCW treatment were noticed, and they were explained, respectively, to understand sulfur removal mechanism (by FR intensity and environment destruction) and metal removal mechanism (by VO2+ complexes’ transformation). For the first time, it was shown that electronic relaxation times extracted from the pulsed EPR measurements can serve as sensitive parameters of SCW treatment. The results confirm that EPR can be used as a complementary tool for analyzing heavy oil upgrading in SCW, even for the online monitoring of oilfield upgrading.

Spin-Resonance Linewidths of Bismuth Donors in Silicon Coupled to Planar Microresonators

Ensembles of bismuth donor spins in silicon are promising storage elements for microwave quantum memories due to their long coherence times which exceed seconds. Operating an efficient quantum memory requires achieving critical coupling between the spin ensemble and a suitable high-quality factor resonator -- this in turn requires a thorough understanding of the lineshapes for the relevant spin resonance transitions, particularly considering the influence of the resonator itself on line broadening. Here, we present pulsed electron spin resonance measurements of ensembles of bismuth donors in natural silicon, above which niobium superconducting resonators have been patterned. By studying spin transitions across a range of frequencies and fields we identify distinct line broadening mechanisms, and in particular those which can be suppressed by operating at magnetic-field-insensitive `clock transitions'. Given the donor concentrations and resonator used here, we measure a cooperativity $C\sim 0.2$ and based on our findings we discuss a route to achieve unit cooperativity, as required for a quantum memory.

Linear and Kinked Oligo(phenyleneethynylene)s as Ideal Molecular Calibrants for Förster Resonance Energy Transfer.

We show that oligo(phenyleneethynylene)s (oligoPEs) are ideal spacers for calibrating dye pairs used for Förster resonance energy transfer (FRET). Ensemble FRET measurements on linear and kinked diads with such spacers show the expected distance and orientation dependence of FRET. Measured FRET efficiencies match excellently with those predicted using a harmonic segmented chain model, which was validated by end-to-end distance distributions obtained from pulsed electron paramagnetic resonance measurements on spin-labeled oligoPEs with comparable label distances.

A Rare Example of Four-Coordinate Nonoxido Vanadium(IV) Alkoxide in the Solid State: Structure, Spectroscopy, and Magnetization Dynamics.

The distorted tetrahedral [V(OAd)4] alkoxide (OAd = 1-adamantoxide, complex 1) is the first homoleptic, mononuclear vanadium(IV) alkoxide to be characterized in the solid state by X-ray diffraction analysis. The compound crystallizes in the cubic P4̅3 n space group with two highly disordered, crystallographically independent molecules in the asymmetric unit. Spin Hamiltonian parameters extracted from low temperature X- and Q-band electron paramagnetic resonance (EPR) experiments performed for polycrystalline samples of 1, both in the concentrated (bulk) form and diluted in the diamagnetic [Ti(OAd)4] analogue, reveal a fully axial system with g z < g x, g y and A z ≫ A x, A y. Complex 1 has also been characterized by alternate current susceptometry with varying temperature (3-30 K) and static magnetic field (up to 8.5 T), showing field-induced slow relaxation of the magnetization with relaxation times ranging from ca. 3 ms at 3 K to 0.02-0.03 ms at 30 K, in line with relevant results described recently for other potential molecular quantum bits. Pulsed EPR measurements, in turn, disclosed long coherence times of ca. 4 μs at temperatures lower than 40 K, despite the presence of the H-rich ligands. The slow spin relaxation in 1 is the first observed for a tetracoordinate nonoxido vanadium(IV) complex, and results are compared here to those generated by square-pyramidal VIV(O)2+ and trigonal prismatic V4+ with oxygen donor atom sets. Considering that the number of promising d1 complexes investigated in detail for slow magnetization dynamics is still small, the present work contributes to the establishment of possible structural/electronic correlations of interest to the field of quantum information processing.

Scaling Up Electronic Spin Qubits into a Three-Dimensional Metal-Organic Framework.

Practical implementation of highly coherent molecular spin qubits for challenging technological applications, such as quantum information processing or quantum sensing, requires precise organization of electronic qubit molecular components into extended frameworks. Realization of spatial control over qubit-qubit distances can be achieved by coordination chemistry approaches through an appropriate choice of the molecular building blocks. However, translating single qubit molecular building units into extended arrays does not guarantee a priori retention of long quantum coherence and spin-lattice relaxation times due to the introduced modifications over qubit-qubit reciprocal distances and molecular crystal lattice phonon structure. In this work, we report the preparation of a three-dimensional (3D) metal-organic framework (MOF) based on vanadyl qubits, [VO(TCPP-Zn2-bpy)] (TCPP = tetracarboxylphenylporphyrinate; bpy = 4,4'-bipyridyl) (1), and the investigation of how such structural modifications influence qubits' performances. This has been done through a multitechnique approach where the structure and properties of a representative molecular building block of formula [VO(TPP)] (TPP = tetraphenylporphyrinate) (2) have been compared with those of the 3D MOF 1. Pulsed electron paramagnetic resonance measurements on magnetically diluted samples in titanyl isostructural analogues revealed that coherence times are retained almost unchanged for 1 with respect to 2 up to room temperature, while the temperature dependence of the spin-lattice relaxation time revealed insights into the role of low-energy vibrations, detected through terahertz spectroscopy, on the spin dynamics.

Spin-Labeling of Proteins via Genetically Encoded Unnatural Amino Acids

Some of the major challenges of protein structure elucidation are proteins that do not crystallize well for x-ray studies and one that give overly convoluted NMR spectra. One way of resolving this issue is through the addition of radical spin-labels for study via electron paramagnetic resonance (EPR). This is usually done through disulfide linkages between a nitroxide spin-label and a cysteine residue of a protein, which allows for continuous wave and pulse EPR measurements. A wealth of information about the system is gained to provide structural insights. However, this requires one to remove native cysteines, which can significantly alter the overall structure and function of the protein. In this investigation, we use the unnatural amino acid, p-azido-phenylalanine, and an alkyne spin probe to “click” our probe to the model protein, T4 lysozyme. This ensures minimal structural perturbation, and allows this method to be used in proteins with native cysteines and disulfide bonds. Through our studies, we observed that this method works, but requires further refinement, and that it is also superior to the cysteine-based method due to the rigidity of the “click” bond, which gives more accurate measurements. Through further study, we hope to expand this method to more complex systems, and streamline a process for quick and easy unnatural protein labeling which will allow researchers to more easily study protein structure and dynamics.

Laser-Induced Magnetic Dipole Spectroscopy.

Pulse electron paramagnetic resonance measurements of nanometer scale distance distributions have proven highly effective in structural studies. They exploit the magnetic dipole-dipole coupling between spin labels site-specifically attached to macromolecules. The most commonly applied technique is double electron-electron resonance (DEER, also called pulsed electron double resonance (PELDOR)). Here we present the new technique of laser-induced magnetic dipole (LaserIMD) spectroscopy based on optical switching of the dipole-dipole coupling. In a proof of concept experiment on a model peptide, we find, already at a low quantum yield of triplet excitation, the same sensitivity for measuring the distance between a porphyrin and a nitroxide label as in a DEER measurement between two nitroxide labels. On the heme protein cytochrome C, we demonstrate that LaserIMD allows for distance measurements between a heme prosthetic group and a nitroxide label, although the heme triplet state is not directly observable by an electron spin echo.

Unexpected suppression of spin-lattice relaxation via high magnetic field in a high-spin iron(iii) complex.

A counterintuitive three-order of magnitude slowing of the spin-lattice relaxation rate is observed in a high spin qubit at high magnetic field via multifrequency pulsed electron paramagnetic resonance measurements.

Distribution and interaction of Mn2+ ions incorporated in cubic ZnS quantum dots over a broad concentration range

The distribution and interaction of isolated Mn2+ impurity ions incorporated in 2.9 nm average diameter cubic ZnS quantum dots (QDs), prepared by surfactant–assisted liquid–liquid synthesis with initial impurity concentrations in the 20–50,000 ppm range, has been investigated by electron paramagnetic resonance (EPR) spectroscopy. The well resolved spectra, observed in the whole investigated concentration range, reflect the localization of the Mn2+ ions in the core and on the surface of the cZnS:Mn QDs at isolated sites. According to the analysis of the dependences of the concentration of incorporated isolated Mn2+ ions vs. initial impurity concentration and of the core localized Mn2+ ions spectra linewidth vs. actual concentration, the isolated Mn2+ ions remain in the whole concentrations range in a diluted paramagnetic state characterized by dipolar magnetic interactions. Pulse EPR measurements of the spin–spin dipolar interaction for the incorporated Mn2+ ions confirm their diluted distribution, which excludes these ions as a possible source of collective magnetism properties.

EPR Study of Structural Phase Transition in Manganese-Doped [(CH3)2NH2][Zn(HCOO)3] Metal–Organic Framework

We present an electron paramagnetic resonance (EPR) study of [(CH3)2NH2][Zn(HCOO)3] metal–organic framework (MOF) powder doped with a small amount of paramagnetic Mn2+ ions. Our EPR measurements indicate a successful incorporation of local Mn2+ probes into the structure allowing us to detect and investigate an order–disorder structural phase transition in the studied MOF. The temperature-dependent continuous wave (CW) X- and Q-band EPR measurements reveal a sudden change in the spectra at a phase transition temperature T0 = 163 K. Simulations were performed to determine the spin Hamiltonian parameters of the spectra which reflect the local symmetry of the Mn2+ probes in the disordered and ordered phases. The temperature dependence of the axial zero-field splitting parameter D demonstrates that the observed phase transition at T0 is discontinuous. Additionally, this dependence follows the prediction of the Landau theory. We also performed preliminary pulse EPR measurements which reveal a rather long phase ...

A high-frequency electron paramagnetic resonance spectrometer for multi-dimensional, multi-frequency, and multi-phase pulsed measurements.

We describe instrumentation for a high-frequency electron paramagnetic resonance (EPR) and pulsed electron-electron double resonance (PELDOR) spectroscopy. The instrumentation is operated in the frequency range of 107-120 GHz and 215-240 GHz and in the magnetic field range of 0-12.1 T. The spectrometer consisting of a high-frequency high-power solid-state source, a quasioptical system, a phase-sensitive detection system, a cryogenic-free superconducting magnet, and a (4)He cryostat enables multi-frequency continuous-wave EPR spectroscopy as well as pulsed EPR measurements with a few hundred nanosecond pulses. Here we discuss the details of the design and the pulsed EPR sensitivity of the instrumentation. We also present performance of the instrumentation in unique experiments including PELDOR spectroscopy to probe correlations in an insulating electronic spin system and application of dynamical decoupling techniques to extend spin coherence of electron spins in an insulating solid-state system.

Redox Modulation of para-Phenylenediamine by Substituted Nitronyl Nitroxide Groups and Their Spin States

Three kinds of para-phenylenediamine (PDA) derivatives bearing nitronyl nitroxide (NN) groups were prepared and characterized on the basis of the electrochemical, electron spin resonance (ESR) spectroscopic, absorption spectroscopic, and magnetic susceptibility measurements. It was clarified that the oxidation potential of the central PDA unit is strongly influenced by the numbers of substituted electron-withdrawing NN groups. In addition, the intervalence charge transfer in the central PDA unit was detected in the monocationic states of the PDAs with two NN groups, indicating the coexistence of the localized spins and the delocalized spin on theses molecules. Moreover, pulsed ESR measurements confirmed that the delocalized spin on the central PDA unit and the localized two spins on the NN groups were ferromagnetically coupled in the monocationic states.

Microwave Loss in the High-Performance DielectricBa(Zn1/3Ta2/3)O3at 4.2 K

Temperature- and magnetic-field-dependent measurements of the loss tangent in Ba(Zn(1/3)Ta(2/3))O(3) doped with transition metals (Mn, Ni) are compared to those from samples doped with other impurities (Cd, Ga, Mg, and Zr). These results, combined with pulsed electron paramagnetic resonance measurements, show conclusively that microwave loss in transition-metal-doped Ba(Zn(1/3)Ta(2/3))O(3) at cryogenic temperatures is attributable to resonant spin excitations of unpaired transition-metal d electrons in isolated atoms (light doping) or exchange coupled clusters (moderate to high doping), a mechanism that differs from the usual suspects.

Electrical activation and electron spin resonance measurements of implanted bismuth in isotopically enriched silicon-28

We have performed continuous wave and pulsed electron spin resonance measurements of implanted bismuth donors in isotopically enriched silicon-28. Donors are electrically activated via thermal annealing with minimal diffusion. Damage from bismuth ion implantation is repaired during thermal annealing as evidenced by narrow spin resonance linewidths (Bpp=12μT) and long spin coherence times (T2=0.7 ms, at temperature T=8 K). The results qualify ion implanted bismuth as a promising candidate for spin qubit integration in silicon.

Nanometer-Range Distance Measurement in a Protein Using Mn2+ Tags

Pulse electron paramagnetic resonance measurements of long-range (nm scale) distances between spin labels site-specifically attached to biomacromolecules have proven highly effective in structural ...