On the peculiar EPR spectra of P1 centers at high (12-20 T) magnetic fields.

The most common lattice defect in high-pressure high-temperature (HPHT) diamonds is the nitrogen substitution (P1) center. This is a paramagnetic defect with a single unpaired electron spin coupled to a N nuclear spin forming an = 1/2, = 1 spin system. While P1 centers have been studied by electron paramagnetic resonance (EPR) spectroscopy for decades, only recently did their behavior at ultra-high (>12 T) magnetic fields become of interest.

High-field pulsed EPR spectroscopy under magic angle spinning.

In this work, we demonstrate the first pulsed electron paramagnetic resonance (EPR) experiments performed under magic angle spinning (MAS) at high magnetic field. Unlike nuclear magnetic resonance (NMR) and dynamic nuclear polarization (DNP), commonly performed at high magnetic fields and under MAS to maximize sensitivity and resolution, EPR is usually measured at low magnetic fields and, with the exception of the Spiess group work in the late 1990s, never under MAS, due to great instrumentational challenges. This hampers the investigation of DNP mechanisms, in which electron spin dynamics play a central role, because no experimental data about the latter under DNP-characteristic conditions are available.

The CW-EPR Capabilities of a Dual DNP/EPR Spectrometer Operating at 14 and 7 T.

High-field electron paramagnetic resonance (EPR) measurements are indispensable for a better understanding of dynamic nuclear polarization (DNP), which relies on polarization transfer between electron and nuclear spins. DNP experiments are typically performed at high > 7 T magnetic fields and low ≤ 100 K temperatures, while EPR instrumentation capable of EPR measurements under these conditions is scarce. In this paper, we describe the CW EPR capabilities of a dual DNP/EPR spectrometer that is designed to carry out EPR experiments under "DNP conditions" at 14 and 7 T.

Nitrogen Substitutions Aggregation and Clustering in Diamonds as Revealed by High-Field Electron Paramagnetic Resonance.

Diamonds have been shown to be an excellent platform for quantum computing and quantum sensing applications. These applications are enabled by the presence of defects in the lattice, which are also known as color centers. The most common nitrogen-based defect in synthetic diamonds is the paramagnetic nitrogen substitution (P1) center.

Carbon-Nitride Popcorn-A Novel Catalyst Prepared by Self-Propagating Combustion of Nitrogen-Rich Triazenes.

The interest in development of non-graphitic polymeric carbon nitrides (PCNs), with various C-to-N ratios, having tunable electronic, optical, and chemical properties is rapidly increasing. Here the first self-propagating combustion synthesis methodology for the facile preparation of novel porous PCN materials (PCN3-PCN7) using new nitrogen-rich triazene-based precursors is reported. This methodology is found to be highly precursor dependent, where variations in the terminal functional groups in the newly designed precursors (compounds 3-7) lead to different combustion behaviors, and morphologies of the resulted PCNs.

Biradical rotamer states tune electron J coupling and MAS dynamic nuclear polarization enhancement.

Cross Effect (CE) Dynamic Nuclear Polarization (DNP) relies on the dipolar (D) and exchange (J) coupling interaction between two electron spins. Until recently only the electron spin D coupling was explicitly included in quantifying the DNP mechanism. Recent literature discusses the potential role of J coupling in DNP, but does not provide an account of the distribution and source of electron spin J coupling of commonly used biradicals in DNP.

Amplification of Dynamic Nuclear Polarization at 200 GHz by Arbitrary Pulse Shaping of the Electron Spin Saturation Profile.

Dynamic nuclear polarization (DNP) takes center stage in nuclear magnetic resonance (NMR) as a tool to amplify its signal by orders of magnitude through the transfer of polarization from electron to nuclear spins. In contrast to modern NMR and electron paramagnetic resonance (EPR) that extensively rely on pulses for spin manipulation in the time domain, the current mainstream DNP technology exclusively relies on monochromatic continuous wave (CW) irradiation. This study introduces arbitrary phase shaped pulses that constitute a train of coherent chirp pulses in the time domain at 200 GHz (7 T) to dramatically enhance the saturation bandwidth and DNP performance compared to CW DNP, yielding up to 500-fold in NMR signal enhancements.

Simple peptide coacervates adapted for rapid pressure-sensitive wet adhesion.

We report here that a dense liquid formed by spontaneous condensation, also known as simple coacervation, of a single mussel foot protein-3S-mimicking peptide exhibits properties critical for underwater adhesion. A structurally homogeneous coacervate is deposited on underwater surfaces as micrometer-thick layers, and, after compression, displays orders of magnitude higher underwater adhesion at 2 N m than that reported from thin films of the most adhesive mussel-foot-derived peptides or their synthetic mimics. The increase in adhesion efficiency does not require nor rely on post-deposition curing or chemical processing, but rather represents an intrinsic physical property of the single-component coacervate.

Thiolate Spin Population of Type I Copper in Azurin Derived from S Hyperfine Coupling.

The electron transfer mediating properties of type I copper proteins stem from the intricate ligand coordination sphere of the Cu ion in their active site. These redox properties are in part due to unusual cysteine thiol coordination, which forms a highly covalent copper-sulfur (Cu-S) bond. The structure and electronic properties of type I copper have been the subject of many experimental and theoretical studies.

Arbitrary waveform modulated pulse EPR at 200GHz.

We report here on the implementation of arbitrary waveform generation (AWG) capabilities at ∼200GHz into an Electron Paramagnetic Resonance (EPR) and Dynamic Nuclear Polarization (DNP) instrument platform operating at 7T. This is achieved with the integration of a 1GHz, 2 channel, digital to analog converter (DAC) board that enables the generation of coherent arbitrary waveforms at K-band frequencies with 1ns resolution into an existing architecture of a solid state amplifier multiplier chain (AMC). This allows for the generation of arbitrary phase- and amplitude-modulated waveforms at 200GHz with >150mW power.

Effect of electron spectral diffusion on static dynamic nuclear polarization at 7 Tesla.

Here, we present an integrated experimental and theoretical study of H dynamic nuclear polarization (DNP) of a frozen aqueous glass containing free radicals at 7 T, under static conditions and at temperatures ranging between 4 and 20 K. The DNP studies were performed with a home-built 200 GHz quasi-optics microwave bridge, powered by a tunable solid-state diode source. DNP using monochromatic and continuous wave (cw) irradiation applied to the electron paramagnetic resonance (EPR) spectrum of the radicals induces the transfer of polarization from the electron spins to the surrounding nuclei of the solvent and solutes in the frozen aqueous glass.

A versatile and modular quasi optics-based 200GHz dual dynamic nuclear polarization and electron paramagnetic resonance instrument.

Solid-state dynamic nuclear polarization (DNP) at higher magnetic fields (>3T) and cryogenic temperatures (∼ 2-90K) has gained enormous interest and seen major technological advances as an NMR signal enhancing technique. Still, the current state of the art DNP operation is not at a state at which sample and freezing conditions can be rationally chosen and the DNP performance predicted a priori, but relies on purely empirical approaches. An important step towards rational optimization of DNP conditions is to have access to DNP instrumental capabilities to diagnose DNP performance and elucidate DNP mechanisms.

Overhauser Dynamic Nuclear Polarization Studies on Local Water Dynamics.

Overhauser dynamic nuclear polarization (ODNP) is an emerging technique for quantifying translational water dynamics in the vicinity (<1 nm) of stable radicals that can be chemically attached to macromolecules of interest. This has led to many in-depth and enlightening studies of hydration water of biomolecules, revolving around the role of solvent dynamics in the structure and function of proteins, nucleic acids, and lipid bilayer membranes. Still to date, a complete and fully automated ODNP instrument is not commercialized.

Distance measurements between manganese(II) and nitroxide spin-labels by DEER determine a binding site of Mn(2+) in the HP92 loop of ribosomal RNA.

Mn(2+) localization in hairpin 92 of the 23S ribosomal RNA (HP92) was obtained using W-band (95 GHz) DEER (double electron-electron resonance) distance measurements between the Mn(2+) ion and nitroxide spin labels on the RNA. It was found to be preferably situated in the minor groove of the double strand region close to the HP92 loop.

Simultaneous DNP enhancements of (1)H and (13)C nuclei: theory and experiments.

DNP on heteronuclear spin systems often results in interesting phenomena such as the polarization enhancement of one nucleus during MW irradiation at the "forbidden" transition frequencies of another nucleus or the polarization transfer between the nuclei without MW irradiation. In this work we discuss the spin dynamics in a four-spin model system of the form {ea-eb-((1)H,(13)C)}, with the Larmor frequencies ωa, ωb, ωH and ωC, by performing Liouville space simulations. This spin system exhibits the common (1)H solid effect (SE), (13)C cross effect (CE) and in addition high order CE-DNP enhancements.

ATPase site configuration of the RNA helicase DbpA probed by ENDOR spectroscopy.

Electron-nuclear double resonance (ENDOR) is a method that probes the local structure of paramagnetic centers via their hyperfine interactions with nearby magnetic nuclei. Here we describe the use of this technique to structurally characterize the ATPase active site of the RNA helicase DbpA, where Mg(2+)-ATP binds. This is achieved by substituting the EPR (electron paramagnetic resonance) silent Mg(2+) ion with paramagnetic, EPR active, Mn(2+) ion.

Determining the oligomeric structure of proteorhodopsin by Gd3+ -based pulsed dipolar spectroscopy of multiple distances.

The structural organization of the functionally relevant, hexameric oligomer of green-absorbing proteorhodopsin (G-PR) was obtained from double electron-electron resonance (DEER) spectroscopy utilizing conventional nitroxide spin labels and recently developed Gd3+ -based spin labels. G-PR with nitroxide or Gd3+ labels was prepared using cysteine mutations at residues Trp58 and Thr177. By combining reliable measurements of multiple interprotein distances in the G-PR hexamer with computer modeling, we obtained a structural model that agrees with the recent crystal structure of the homologous blue-absorbing PR (B-PR) hexamer.

Correlating nuclear frequencies by two-dimensional ELDOR-detected NMR spectroscopy.

ELDOR (Electron Double Resonance)-detected NMR (EDNMR) is a pulse EPR experiment that is used to measure the transition frequencies of nuclear spins coupled to electron spins. These frequencies are further used to determine hyperfine and quadrupolar couplings, which are signatures of the electronic and spatial structures of paramagnetic centers. In recent years, EDNMR has been shown to be particularly useful at high fields/high frequencies, such as W-band (∼95 GHz, ∼3.

W-band orientation selective DEER measurements on a Gd3+/nitroxide mixed-labeled protein dimer with a dual mode cavity.

Double electron-electron resonance (DEER) at W-band (95 GHz) was applied to measure the distance between a pair of nitroxide and Gd(3+) chelate spin labels, about 6 nm apart, in a homodimer of the protein ERp29. While high-field DEER measurements on systems with such mixed labels can be highly attractive in terms of sensitivity and the potential to access long distances, a major difficulty arises from the large frequency spacing (about 700 MHz) between the narrow, intense signal of the Gd(3+) central transition and the nitroxide signal. This is particularly problematic when using standard single-mode cavities.

Topology of the trans-membrane peptide WALP23 in model membranes under negative mismatch conditions.

The organization and orientation of membrane-inserted helices is important for better understanding the mode of action of membrane-active peptides and of protein-membrane interactions. Here we report on the application of ESEEM (electron spin-echo envelope modulation) and DEER (double electron-electron resonance) techniques to probe the orientation and oligomeric state of an α-helical trans-membrane model peptide, WALP23, under conditions of negative mismatch between the hydrophobic cores of the model membrane and the peptide. Using ESEEM, we measured weak dipolar interactions between spin-labeled WALP23 and (2)H nuclei of either the solvent (D2O) or of lipids specifically deuterated at the choline group.

Spectroscopic selection of distance measurements in a protein dimer with mixed nitroxide and Gd3+ spin labels.

The pulse DEER (Double Electron-Electron Resonance) technique is frequently applied for measuring nanometer distances between specific sites in biological macromolecules. In this work we extend the applicability of this method to high field distance measurements in a protein assembly with mixed spin labels, i.e.

Probing conformational variations at the ATPase site of the RNA helicase DbpA by high-field electron-nuclear double resonance spectroscopy.

The RNA helicase DbpA promotes RNA remodeling coupled to ATP hydrolysis. It is unique because of its specificity to hairpin 92 of 23S rRNA (HP92). Although DbpA kinetic pathways leading to ATP hydrolysis and RNA unwinding have been recently elucidated, the molecular (atomic) basis for the coupling of ATP hydrolysis to RNA remodeling remains unclear.

W-Band pulse EPR distance measurements in peptides using Gd(3+)-dipicolinic acid derivatives as spin labels.

We present high field DEER (double electron-electron resonance) distance measurements using Gd(3+) (S = 7/2) spin labels for probing peptides' conformations in solution. The motivation for using Gd(3+) spin labels as an alternative for the standard nitroxide spin labels is the sensitivity improvement they offer because of their very intense EPR signal at high magnetic fields. Gd(3+) was coordinated by dipicolinic acid derivative (4MMDPA) tags that were covalently attached to two cysteine thiol groups.