MR imaging with phase encoding of intermolecular multiple quantum coherences

Novel 2D and 3D pulse sequences producing images through the phase encoding of intermolecular multiple quantum coherences (i-MQCs) are presented. The signal acquired with these sequences is free from intermolecular zero quantum coherences (i-ZQCs) which are not phase encoded and additional phase cycling eliminates artifacts. Phase encoding during the n-quantum evolution period provides n times the resolution expected from equivalent phase encoding of the reconverted single quantum coherences. These sequences have potential applications for producing i-MQC images of biological tissues as well as nonbiological materials with substantial amounts of water.     

Simplified multiplet pattern HSQC-TOCSY experiment for accurate determination of long-range heteronuclear coupling constants

A new two-dimensional pulse sequence for accurately determining heteronuclear coupling constants is presented. It is derived from HSQC and HECADE techniques with B0 gradient coherence selection. The main feature of the proposed method is spectra with only one component of the IS doublet; i.e., the final result is equivalent to a selective broadband excitation of either Salpha or Sbeta spin states and a preservation of these states during the entire experiment. The effect is obtained by an appropriate combination of in- and antiphase coherences. It is demonstrated that heteronuclear single-bond as well as long-range coupling constants and their relative signs are readily evaluated. The proposed sequence is equally or less sensitive to a variation of heteronuclear one-bond couplings than previously published, closely related sequences. The new method is applied to a peptide sample for determination of 3JN, Hbeta.     

Novel methods for characterizing a decoupler channel using "undetectable" quantum coherences.

Exact solutions for the effect of time-independent RF pulses on any initial configuration of an IS J-coupled system demonstrate that on-resonance CW decoupling yields signals whose frequency depends on RF field strength and homogeneity. These signals are enhanced starting with "undetectable" antiphase and multiple quantum coherences, which can also produce centerband intensity to mimic the signal from decoupled Sx. Conversely, these coherences can be generated from Sx using a low-power pulse, B1 = J/2, of length (2J)-1, dubbed a "90J pulse" since it is the selective equivalent of {(2J)-1-90[I]}. Utilizing 90J pulses, new characterization-of-decoupler (COD) pulse sequences can determine the performance of an insensitive I-spin channel by observing large signals from either antiphase or multiple quantum coherences with the S-spin channel, allowing, in minutes rather than hours: (i) frequency calibration to an accuracy of 0.1 Hz; (ii) measurement of RF amplitudes over a 500-fold variation; and (iii) mapping of RF homogeneity along the sample axis with a single 1D B1 spectrum. These 90J coherence transfer pulses are of potential general use for selective spectroscopy.     

Homo- and heteronuclear 2D NMR approaches to analyse a mixture of deuterated unlike/like stereoisomers using weakly ordering chiral liquid crystals

We describe several homo- and heteronuclear 2D NMR strategies dedicated to the analysis of anisotropic (2)H spectra of a mixture of dideuterated unlike/like stereoisomers with two remote stereogenic centers, using weakly orienting chiral liquid crystals. To this end, we propose various 2D correlation experiments, denoted "D(H)(n)D" or "D(H)(n)C" (with n=1, 2), that involve two heteronuclear polarization transfers of INEPT-type with one or two proton relays. The analytical expressions of correlation signals for four pulse sequences reported here were calculated using the product-operators formalism for spin I=1 and S=1/2. The features and advantages of each scheme are presented and discussed. The efficiency of these 2D sequences is illustrated using various deuterated model molecules, dissolved in organic solutions of polypeptides made of poly-gamma-benzyl-L-glutamate (PBLG) or poly-epsilon-carbobenzyloxy-L-lysine (PCBLL) and NMR numerical simulations.     

多维核磁共振波谱学

核磁共振波谱学自本世纪40年代中期诞生起,已走完了其光辉的50年历程,其间,它已完成了自连续波检测向脉冲傅里叶变换检测的过渡,并在此基础上率先将传统的一维谱的观测拓宽为多维谱的观测,不断提供了其它方法难以提供或无法提供的崭新信息,从而确立了它在分子结构和动力学研究中的独特地位,本文旨在以二维NMR为主,简述被称为现代核磁共振波谱学之核心的多维NMK的概况。     

Optimization of Three-Dimensional TROSY-Type HCCH NMR Correlation of Aromatic H–C Groups in Proteins

Improved methods for three-dimensional TROSY-Type HCCH correlation involving protons of negligible CSA are presented. The TROSY approach differs from the conventional approach of heteronuclear decoupling in evolution and detection periods by not mixing fast and slowly relaxing coherences and usually suppressing the former. Pervushin et al. (J. Am. Chem. Soc. 120, 6394–6400 (1998)) have proposed a 3D TROSY-type HCCH experiment where the TROSY approach is applied only in one of the ¹³C dimensions. A new pulse sequence applying the TROSY approach in both indirect dimensions is advantageous when the TROSY effect of the carbons is large or when a relatively high resolution is required. For lower resolutions or moderate TROSY effects we show that it is possible to combine the best of both worlds, namely to suppress heteronuclear couplings without mixing fast and slowly relaxing coherences while at the same time superimpose the two components and thus have both contribute to the detected signal. That is possible using the novel technique of Spin-State-Selective Time-Proportional Phase Incrementation (S³ TPPI). The new 3D S³ TPPI TROSY HCCH method is demonstrated on a ¹³C,¹⁵N-labeled protein sample, RAP 18–112 (N-terminal domain of α₂-macroglobulin receptor associated protein), at 750 MHz and average sensitivity enhancements of 10% are obtained for the cross peaks in comparison to methods based on conventional decoupling on one of the carbons or on TROSY on both carbons.     

Heteronuclear intermolecular single-quantum coherences in liquid nuclear magnetic resonance

This paper analyses the heteronuclear Cosy Revamped by Asymmetric Z-gradient Echo Detection pulse sequence. General theoretical expressions of the pulse sequence with arbitrary flip angles were derived by using dipolar field treatment and signals originating from heteronuclear intermolecular single-quantum coherences （iSQCs） in highly-polarized two spin-1/2 systems were mainly discussed in order to find the optimal flip angles. The results show that signals from heteronuclear iSQCs decay slower than those from intermolecular double-quantum coherences or intermolecular zero-quantum coherences. Magical angle experiments validate that the signals are from heteronuclear iSQCs and insensitive to the imperfection of radio-frequency flip angles. All experimental observations are in excellent agreement with theoretical predictions. The quantum-mechanical treatment leads to similar predictions to the dipolar field treatment.     

Inhomogeneity-free heteronuclear iMQC

Intermolecular dipolar interactions between proton and carbon spins can be used to indirectly detect carbon spectra with high sensitivity. In this communication, we present a modified sequence that, in addition to the high sensitivity of heteronuclear intermolecular multiple quantum coherence (iMQC) experiments, retains the line narrowing capability characteristic of homonuclear zero-quantum coherences. We demonstrate that this sequence can be used to obtain high resolution (13)C spectra in the presence of magnetic field inhomogeneities, both for thermal and hyperpolarized samples, and discuss applications to water-hyperpolarized carbon imaging.     

Selection rules for multiple quantum NMR excitation in solids: derivation from time-reversal symmetry and comparison with simulations and (13)C NMR experiments.

New derivations of selection rules for excitation and detection of multiple quantum coherences in coupled spin-1/2 systems are presented. The selection rules apply to experiments in which the effective coupling Hamiltonian used for multiple quantum excitation is both time-reversal invariant and time-reversible by a phase shift of the radiofrequency pulse sequence that generates the effective couplings. The selection rules are shown to be consequences of time-reversal invariance and time-reversibility and otherwise independent of the specific form of the effective coupling Hamiltonian. Numerical simulations of multiple quantum NMR signal amplitudes and experimental multiple quantum excitation spectra are presented for the case of a multiply (13)C-labeled helical polypeptide. The simulations and experiments confirm the selection rules and demonstrate their impact on multiple quantum (13)C NMR spectra in this biochemically relevant case.     

High-resolution 2D NMR spectra in inhomogeneous fields based on intermolecular multiple-quantum coherences with efficient acquisition schemes

High-resolution 2D NMR spectra in inhomogeneous fields can be achieved by the use of intermolecular multiple-quantum coherences and shearing reconstruction of 3D data. However, the long acquisition time of 3D spectral data is generally unbearable for in vivo applications. To overcome this problem, two pulse sequences dubbed as iDH-COSY and iDH-JRES were proposed in this paper. Although 3D acquisition is still required for the new sequences, the high-resolution 2D spectra can be obtained with a relatively short scanning time utilizing the manipulation of indirect evolution period and sparse sampling. The intermolecular multiple-quantum coherence treatment combined with the raising and lowering operators was applied to derive analytical signal expressions for the new sequences. And the experimental observations agree with the theoretical predictions. Our results show that the new sequences possess bright perspective in the applications on in vivo localized NMR spectroscopy.     

13C PGSE NMR experiment with heteronuclear dipolar decoupling to measure diffusion in liquid crystals and solids

A new PGSE NMR experiment, designed to measure molecular diffusion coefficients in systems with nonvanishing static dipolar coupling, is described. The fast static dipolar dephasing of the single-quantum (13)C coherences is removed by multiple-pulse heteronuclear decoupling. The resulting slow dephasing of the (13)C coherences allows for inserting appropriate gradient pulses into the pulse sequence. The presence of the large magnetic field gradient reduces the efficiency of the decoupling sequences which is compensated for by introducing a scheme of sequential slice selection across the sample. The method is demonstrated by (19)F-decoupled (13)C PGSE NMR experiments in a lyotropic nematic and lamellar liquid crystal.     

Optimization of three-dimensional TROSY-type HCCH NMR correlation of aromatic (1)H-(13)C groups in proteins.

Improved methods for three-dimensional TROSY-Type HCCH correlation involving protons of negligible CSA are presented. The TROSY approach differs from the conventional approach of heteronuclear decoupling in evolution and detection periods by not mixing fast and slowly relaxing coherences and usually suppressing the former. Pervushin et al. (J. Am. Chem. Soc. 120, 6394-6400 (1998)) have proposed a 3D TROSY-type HCCH experiment where the TROSY approach is applied only in one of the (13)C dimensions. A new pulse sequence applying the TROSY approach in both indirect dimensions is advantageous when the TROSY effect of the carbons is large or when a relatively high resolution is required. For lower resolutions or moderate TROSY effects we show that it is possible to combine the best of both worlds, namely to suppress heteronuclear couplings without mixing fast and slowly relaxing coherences while at the same time superimpose the two components and thus have both contribute to the detected signal. That is possible using the novel technique of Spin-State-Selective Time-Proportional Phase Incrementation (S(3) TPPI). The new 3D S(3) TPPI TROSY HCCH method is demonstrated on a (13)C,(15)N-labeled protein sample, RAP 18-112 (N-terminal domain of alpha(2)-macroglobulin receptor associated protein), at 750 MHz and average sensitivity enhancements of 10% are obtained for the cross peaks in comparison to methods based on conventional decoupling on one of the carbons or on TROSY on both carbons.     

2H T2rho relaxation dynamics and double-quantum filtered NMR studies

In this study 2H T2rho DQF NMR spectra of water in MCM-41 were measured. The T2rho double-quantum filtered (DQF) NMR signal is generated by applying a radio frequency (RF) field for various durations and then observed after a monitor RF pulse. It was found that the transfer between different quantum coherences by the couplings during long-duration RF fields (i.e., soft pulses) and that residual quadrupolar interaction dominates the signal decay. Knowledge of coherence transfer during long-RF pulses has special significance for the development of sophisticated multi-quantum NMR experiments especially multi-quantum MRI applications.     

Numerical simulation of one- and two-dimensional ESEEM experiments

Numerical simulation has become an indispensable tool for the interpretation of pulse EPR experiments. In this work it is shown how automatic orientation selection, grouping of operator factors, and direct selection and elimination of coherences can be used to improve the efficiency of time-domain simulations of one- and two-dimensional electron spin echo envelope modulation (ESEEM) spectra. The program allows for the computation of magnetic interactions of any symmetry and can be used to simulate spin systems with an arbitrary number of nuclei with any spin quantum number. Experimental restrictions due to finite microwave pulse lengths are addressed and the enhancement of forbidden coherences by microwave pulse matching is illustrated. A comparison of simulated and experimental HYSCORE (hyperfine sublevel correlation) spectra of ordered and disordered systems with varying complexity shows good qualitative agreement.     

Time-optimal coherence-order-selective transfer of in-phase coherence in heteronuclear IS spin systems

Based on principles of geometric optimal control theory, coherence transfer building blocks can be derived which achieve optimal sensitivity. Here, experimental pulse sequences are presented that achieve the best possible coherence-order-selective in-phase transfer (S(-)-->I(-)) for a heteronuclear 2-spin system for any given mixing time in the absence of relaxation. For short mixing times, the optimal experiment improves the sensitivity of isotropic mixing by up to 12.5%.     

A new sequence for the MQMAS/HETCOR experiment

Two-dimensional (2D) multiple quantum MAS (magic angle spinning) spectroscopy has been combined with cross-polarisation to obtain a heteronuclear correlation spectrum between a quadrupolar spin-3/2 and a spin-1/2 nucleus. The advantage over the conventional correlation experiment is the increased resolution obtained in the multiple quantum dimension. Pure absorption 2D spectra can be obtained by implementing a zero quantum filter between the evolution of multiple quanta and the subsequent cross-polarisation step. The current experiment shows a considerable improvement in sensitivity compared to a previously introduced sequence.     

Selective Correlation of Amide Groups to Glycine Alpha Protons and of Arginine Guanidine Groups to Delta Protons in Proteins by Multiple Quantum Spectroscopy

A new 3D pulse sequence correlates backbone amide proton and nitrogen with alpha proton resonances selectively for glycine residues in a fully doubly labeled (¹⁵N,¹³C) protein. The excitation of multiple quantum coherences provides optimized resolution and sensitivity. Degenerate alpha proton groups can be promptly recognized. Correlation of guanidine NH groups to delta protons of arginine side chains is also obtained.     

Selection Rules for Multiple Quantum NMR Excitation in Solids: Derivation from Time-Reversal Symmetry and Comparison with Simulations and C NMR Experiments

New derivations of selection rules for excitation and detection of multiple quantum coherences in coupled spin-1/2 systems are presented. The selection rules apply to experiments in which the effective coupling Hamiltonian used for multiple quantum excitation is both time-reversal invariant and time-reversible by a phase shift of the radiofrequency pulse sequence that generates the effective couplings. The selection rules are shown to be consequences of time-reversal invariance and time-reversibility and otherwise independent of the specific form of the effective coupling Hamiltonian. Numerical simulations of multiple quantum NMR signal amplitudes and experimental multiple quantum excitation spectra are presented for the case of a multiply ¹³C-labeled helical polypeptide. The simulations and experiments confirm the selection rules and demonstrate their impact on multiple quantum ¹³C NMR spectra in this biochemically relevant case.     

Heteronuclear Correlation Spectroscopy with RF Gradients

The applicability of RF gradients for suppressing the resonances of uncoupled spins in inverse-detected heteronuclear spectroscopy has been investigated. Pulse sequences were designed which incorporate RF gradients in heteronuclear single-quantum-correlation experiments (HSQC), and they can be divided into three categories based on how the RF gradients are used. In the first type of experiment, the desired coherences are spin locked in the RF-gradient held, while unwanted terms, placed perpendicular to the direction of the RF-gradient held, are dephased. In a second type of experiment, the dephasing action of the gradient and the coherence-transfer RF pulses are combined into a single RF-gradient pulse. A second RF-gradient pulse is then used to rephase the desired spin terms. The third type of experiment uses a period of longitudinal storage of the heteronuclear magnetization, during which time the magnetization of the uncoupled spins is destroyed by an RF-gradient pulse. Experimental results are shown from all three techniques, and the techniques are compared.     

HMSC: simultaneously detected heteronuclear shift correlation through multiple and single bonds

A new 2D pulse sequence HMSC (heteronuclear multiple-bond and single-bond coupling connectivities) for the simultaneous detection of long-range and one-bond heteronuclear connectivities is proposed which allows the two types of responses to be separated and the corresponding (n)J(CH) and (1)J(CH) connectivity maps to be calculated. (n)J(CH) coherences are selectively labeled in the course of the pulse sequence, the correspondingly acquired data are separately stored, and a simple add/subtract procedure is applied to disentangle and edit (n)J(CH) and (1)J(CH) responses prior to final data processing. Unlike standard methods, which are designed to measure one single type of heteronuclear spin-spin interactions and to efficiently suppress the other, both (n)J(CH) and (1)J(CH) are measured simultaneously in a single experiment with the HMSC pulse sequence. Compared to the common strategy with two standard experiments applied one after the other, e.g., HMBC and HMQC, valuable measuring time may be saved with this single experiment approach. The efficiency of the new pulse sequence and the quality of the corresponding spectra are demonstrated using strychnine. Features such as sensitivity, lineshapes, and the suppression of (1)J(CH) residual peaks in the final (n)J(CH) subspectra are investigated and compared with the corresponding results obtained with standard methods. The attractive and unique single experiment approach, its high efficiency, and its easy experimental setup together with straightforward data processing make HMSC a valuable experimental alternative for the today's more time-consuming "two-step" practice and makes it suitable for standard routine applications.