Improved detection of magnetic interactions in proteins based on long-lived coherences.

Living systems rely on molecular building blocks with low structural symmetry. Therefore, constituent amino acids and nucleotides yield short-lived nuclear magnetic responses to electromagnetic radiation. Magnetic signals are at the basis of molecular imaging, structure determination and interaction studies.

Cross polarization from dipolar-order under magic angle spinning: The ADRF-CPMAS NMR experiment.

Techniques for enhancing the signals arising from low-γ, insensitive (I) nuclei are central to solid-state nuclear magnetic resonance. One of the leading and best-established methods to sensitize these unreceptive species is Hartmann-Hahn cross polarization (HH-CP), a polarization transfer mechanism often executed under MAS. Herein, we explore the possibility of utilizing the 1H dipolar order created via adiabatic demagnetization in the rotating frame (ADRF), to enhance the unreceptive spins under MAS.

Identifying and Overcoming Artifacts in H-Based Saturation Transfer NOE NMR Experiments.

Magnetization transfer experiments are versatile nuclear magnetic resonance (NMR) tools providing site-specific information. We have recently discussed how saturation magnetization transfer (SMT) experiments could leverage repeated repolarizations arising from exchanges between labile and water protons to enhance connectivities revealed via the nuclear Overhauser effect (NOE). Repeated experience with SMT has shown that a number of artifacts may arise in these experiments, which may confound the information being sought - particularly when seeking small NOEs among closely spaced resonances.

Effective Hamiltonian and spin dynamics in fast MAS TRAPDOR-HMQC experiments involving spin-3/2 quadrupolar nuclei.

We present a theoretical and numerical description of the spin dynamics associated with TRAPDOR-HMQC (T-HMQC) experiment for a H (I) - Cl (S) spin system under fast magic angle spinning (MAS). Towards this an exact effective Hamiltonian describing the system is numerically evaluated with matrix logarithm approach. The different magnitudes of the heteronuclear and pure S terms in the effective Hamiltonian allow us to suggest a truncation approximation, which is shown to be in excellent agreement with the exact time evolution.

Selective Excitation of Long-Lived Nuclear Spin States.

Nuclear magnetization storage, once limited by longitudinal and transverse relaxation lifetimes, and , can be prolonged by symmetry-adapted nuclear spin order, i.e. long-lived states (LLS) and long-lived coherences (LLC), which have significantly extended relaxation time constants compared to and , respectively.

The Extended Hadamard Transform: Sensitivity-Enhanced NMR Experiments Among Labile and Non-Labile Hs of SARS-CoV-2-derived RNAs.

Hadamard encoded saturation transfer can significantly improve the efficiency of NOE-based NMR correlations from labile protons in proteins, glycans and RNAs, increasing the sensitivity of cross-peaks by an order of magnitude and shortening experimental times by ≥100-fold. These schemes, however, fail when tackling correlations within a pool of labile protons - for instance imino-imino correlations in RNAs or amide-amide correlations in proteins. Here we analyze the origin of the artifacts appearing in these experiments and propose a way to obtain artifact-free correlations both within the labile pool as well as between labile and non-labile Hs, while still enjoying the gains arising from Hadamard encoding and solvent repolarizations.

Heteronuclear transfers from labile protons in biomolecular NMR: Cross polarization, revisited.

INEPT- and HMQC-based pulse sequences are widely used to transfer polarization between heteronuclei, particularly in biomolecular spectroscopy: they are easy to setup and involve low power deposition. Still, these short-pulse polarization transfers schemes are challenged by fast solvent chemical exchange. An alternative to improve these heteronuclear transfers is J-driven cross polarization (J-CP), which transfers polarization by spin-locking the coupled spins under Hartmann-Hahn conditions.

3D Heteronuclear Magnetization Transfers for the Establishment of Secondary Structures in SARS-CoV-2-Derived RNAs.

Multidimensional NOESY experiments targeting correlations between exchangeable imino and amino protons provide valuable information about base pairing in nucleic acids. It has been recently shown that the sensitivity of homonuclear correlations involving RNA's labile imino protons can be significantly enhanced, by exploiting the repolarization brought about by solvent exchanges. Homonuclear correlations, however, are of limited spectral resolution, and usually incapable of tackling relatively large homopolymers with repeating structures like RNAs.

Effective Hamiltonian and H-N cross polarization/double cross polarization at fast MAS.

In this work we investigate in detail the underlying spin-dynamics associated with H-N double CP experiments under fast MAS, recently demonstrated by Carnevale et al. We employ matrix logarithm and Floquet theory to compute numerically the effective Hamiltonian associated to the time-dependent problem. Certain common features related to construction of effective Hamiltonians by both approaches are discussed.

Sensitivity enhancement in 2D Double Cross Polarization experiments under fast MAS by recycling unused protons.

We demonstrate sensitivity enhancement via recycling of proton magnetization in 2D Double Cross Polarization (Double CP) experiments performed on fully protonated and uniformly labeled (C, N) samples at a magic angle spinning rate of 60 kHz. Unused proton magnetization is preserved during t evolution either by locking it with CW irradiation or by employing rotor-synchronized pi pulses. A flip-back pulse together with a modified second CP block preserves unused proton magnetization resulting in enhanced sensitivity.

Multidimensional J-driven NMR correlations by single-scan offset-encoded recoupling.

Two-dimensional (2D) correlations between bonded heteroatoms, lie at the cornerstone of many uses given to contemporary nuclear magnetic resonance (NMR). Improving the efficiency with which these correlations are established is an important topic in modern NMR, with potential applications in rapid chemical analysis and dynamic biophysical studies. Alternatives have been developed over the last decade to speed up these experiments, based among others on reducing the number of data points that need to be sampled, and/or shortening the inter-scan delays.

Heteronuclear cross-relaxation effects in the NMR spectroscopy of hyperpolarized targets.

Dissolution dynamic nuclear polarization (DNP) enables high-sensitivity solution-phase NMR experiments on long-lived nuclear spin species such as (15)N and (13)C. This report explores certain features arising in solution-state (1)H NMR upon polarizing low-γ nuclear species. Following solid-state hyperpolarization of both (13)C and (1)H, solution-phase (1)H NMR experiments on dissolved samples revealed transient effects, whereby peaks arising from protons bonded to the naturally occurring (13)C nuclei appeared larger than the typically dominant (12)C-bonded (1)H resonances.

Rapid acquisition of 14N solid-state NMR spectra with broadband cross polarization.

Nitrogen is an element of utmost importance in chemistry, biology and materials science. Of its two NMR-active isotopes, (14)N and (15)N, solid-state NMR (SSNMR) experiments are rarely conducted upon the former, due to its low gyromagnetic ratio (γ) and broad powder patterns arising from first-order quadrupolar interactions. In this work, we propose a methodology for the rapid acquisition of high quality (14)N SSNMR spectra that is easy to implement, and can be used for a variety of nitrogen-containing systems.

Two-dimensional RF pulses: a new approach to selectively exciting J-coupled spins in nuclear magnetic resonance.

A new scheme for the excitation of spins according to the joint values of their heteronuclear or homonuclear J couplings and of their chemical shifts, is proposed and demonstrated. The principles of the new pulses involved derive from those employed in NMR imaging for exciting arbitrary 2D spatial shapes, using so-called "multidimensional" RF pulses. It is shown that if recast in a suitable spectroscopic framework, the distinction that π-pulses enable to establish between linear and bilinear interactions, support the selective excitation of coherences possessing arbitrary combinations of chemical shift and J-coupling values--a flexibility akin to that provided by a 2D J-resolved NMR spectrum.

Broadband adiabatic inversion pulses for cross polarization in wideline solid-state NMR spectroscopy.

Efficient acquisition of ultra-wideline solid-state NMR powder patterns is a continuing challenge. In particular, when the breadth of the powder pattern is much larger than the cross-polarization (CP) excitation bandwidth, transfer efficiencies suffer and experimental times are greatly increased. Presented herein is a CP pulse sequence with an excitation bandwidth that is up to ten times greater than that available from a conventional spin-locked CP pulse sequence.

Toward single-shot pure-shift solution 1H NMR by trains of BIRD-based homonuclear decoupling.

Achieving homonuclear 1H decoupling remains one of the key challenges in liquid-state NMR. Such spectra would endow a variety of organic and analytical applications with an increased resolution, and would ideally do so even in a one-dimensional format. A number of parallel efforts aimed at achieving this goal using two-dimensional acquisitions have been proposed; approaches demonstrated over recent years include, among others, new modes for achieving purely-absorptive J spectroscopy, the use of spatially-selective manipulations, and exploiting the natural spin dilution afforded by heteronuclei.

Sensitizing solid state nuclear magnetic resonance of dilute nuclei by spin-diffusion assisted polarization transfers.

Recent years have witnessed efforts geared at increasing the sensitivity of NMR experiments, by relying on the suitable tailoring and exploitation of relaxation phenomena. These efforts have included the use of paramagnetic agents, enhanced (1)H-(1)H incoherent and coherent transfers processes in 2D liquid state spectroscopy, and homonuclear (13)C-(13)C spin diffusion effects in labeled solids. The present study examines some of the opportunities that could open when exploiting spontaneous (1)H-(1)H spin-diffusion processes, to enhance relaxation and to improve the sensitivity of dilute nuclei in solid state NMR measurements.

Indirect detection of nitrogen-14 in solid-state NMR spectroscopy.

NMR spectra of (14)N (spin I=1) are obtained by indirect detection in powders spinning at the magic angle. The method relies on the transfer of coherence from a neighboring "spy" nucleus with S=1/2, such as (13)C or (1)H, to single- or double-quantum transitions of (14)N nuclei. The transfer of coherence can occur through a combination of scalar and residual dipolar splittings (RDS); the latter are also known as second-order quadrupole-dipole cross terms.

Indirect detection of nitrogen-14 in solids via protons by nuclear magnetic resonance spectroscopy.

This Communication describes the indirect detection of 14N nuclei (spin I=1) in solids by nuclear magnetic resonance (NMR) spectroscopy. The two-dimensional correlation method used here is closely related to the heteronuclear multiple quantum correlation (HMQC) experiment introduced in 1979 to study molecules in liquids, which has recently been used to study solids spinning at the magic angle. The difference is that the coherence transfer from neighboring 1H nuclei to 14N is achieved via a combination of J couplings and residual dipolar splittings (RDS).

Nitrogen-14 NMR spectroscopy using residual dipolar splittings in solids.

It is shown that nuclear magnetic resonance (NMR) spectra of nitrogen-14 (spin I = 1) can be obtained by indirect detection in powders spinning at the magic angle (MAS). The method relies on the transfer of coherence from a neighboring nucleus with S = 1/2, such as carbon-13, to single- or double-quantum transitions of nitrogen-14 nuclei. The transfer of coherence occurs through second-order quadrupole-dipole cross terms, also known as residual dipolar splittings.

Principles and features of single-scan two-dimensional NMR spectroscopy.

Two-dimensional nuclear magnetic resonance (2D NMR) provides one of the foremost contemporary tools available for the elucidation of molecular structure, function, and dynamics. Execution of a 2D NMR experiment generally involves scanning a series of time-domain signals S(t(2)), as a function of a t(1) time variable which undergoes parametric incrementation throughout independent experiments. Very recently, we proposed and demonstrated a general approach whereby this serial mode of data acquisition is parallelized, enabling the acquisition of complete bidimensional NMR data sets via the recording of a single transient.

Line shape considerations in ultrafast 2D NMR.

We have recently proposed and demonstrated an approach that enables the acquisition of 2D nuclear magnetic resonance (NMR) spectra within a single scan. The approach is based on spatially encoding the spins' evolution along the indirect domain with the aid of a magnetic field gradient, and subsequently decoding this information numerous times over the course of the signal acquisition while spins are subject to a train of gradient echoes. The present paper discusses further considerations pertaining the 2D line shapes arising from this new way of collecting NMR data.

Relaxation-assisted separation of chemical sites in NMR spectroscopy of static solids.

We discuss the potential use of relaxation times toward the resolution of inequivalent chemical sites in the NMR spectroscopy of powdered or disordered samples. This proposal is motivated by the significant differences that can often be detected in the relaxation behavior of sites in solids, particularly when focusing on NMR observations of quadrupolar nuclei possessing different coordination and/or dynamic environments. It is shown that in these cases the implementation of a non-negative least-squares analysis on relaxation data sets enables the bidimensional resolution of overlapping powder line shapes, even when dealing with static samples.

The acquisition of multidimensional NMR spectra within a single scan.

A scheme enabling the complete sampling of multidimensional NMR domains within a single continuous acquisition is introduced and exemplified. Provided that an analyte's signal is sufficiently strong, the acquisition time of multidimensional NMR experiments can thus be shortened by orders of magnitude. This could enable the characterization of transient events such as proteins folding, 2D NMR experiments on samples being chromatographed, bring the duration of higher dimensional experiments (e.

Rotor-encoded heteronuclear MQ MAS NMR spectroscopy of half-integer quadrupolar and spin I=1/2 nuclei.

A new two-dimensional heteronuclear multiple-quantum magic-angle spinning (MQ MAS) experiment is presented which combines high resolution for the half-integer quadrupolar nucleus with information about the dipolar coupling between the quadrupolar nucleus and a spin I=1/2 nucleus. Homonuclear MQ coherence is initially created for the half-integer quadrupolar nucleus by a single pulse as in a standard MQ MAS experiment. REDOR recoupling of the heteronuclear dipolar coupling then allows the creation of a heteronuclear multiple-quantum coherence comprising multiple- and single-quantum coherence of the quadrupolar and spin I=1/2 nucleus, respectively, which evolves during t1.