固体核磁共振中膜蛋白双交叉极化效率与动力学参数相关的定量分析

固体核磁共振（NMR）中双交叉极化（DCP）是用于膜蛋白信号指认的多维异核相关实验的基本技术模块.DCP的效率在很大程度上决定了多维异核相关实验的效率.本文分析了3种典型的膜环境中的膜蛋白（AQPZ、DAGK和EV71 2B）的DCP效率及其影响因素.结果显示,在相同的实验条件下,3种蛋白样品的DCP效率存在明显差异：其中AQPZ的DCP效率最高（31%）,DAGK的效率次之（23%）,EV71 2B的效率最低（14%）.通过测量它们在旋转坐标下的自旋-晶格弛豫时间（T_1ρ）和偶极耦合常数（D_HN）,发现膜蛋白的运动会明显缩短T_1ρ,但对D_HN的影响较小.在实验的基础上,建立了T_1ρ与DCP效率相关的模型,并基于DCP动力学的定量分析,证明了运动导致的T_1ρ缩短是降低DCP效率的主要原因.因此,可以通过定量分析未知样品的T_1ρ来预测其DCP的最优效率,为DCP实验的优化提供依据.     

生物固体核磁共振中样品发热的研究进展

近年来,固体核磁共振被广泛应用于膜蛋白、纤维化蛋白等体系的结构和功能研究．在固体核磁共振实验中,快速魔角旋转或高功率射频场照射等实验条件将导致样品发热．生物样品发热能导致严重的后果,例如样品温度的快速升高,信号分辨率、信噪比的降低,发热严重时甚至导致样品的不可逆损坏．近年来,人们对样品发热问题进行了一些研究,发现通过优化样品制备条件或固体核磁共振实验条件,以及改进探头设计等手段,可以在一定程度上减轻样品发热．该文主要综述了生物固体核磁共振研究中导致样品发热的原因和减轻样品发热的方法．     

具有快速调制功能NMR发射机的设计与实现

介绍了一种具有快速幅度、相位和频率调制功能的核磁共振射频发射机. 数字中频子系统中采用了两个级联的数控振荡器,前一级数控振荡器在FPGA内实现,用于产生基带调制信号;后一级则采用具有正交调制功能的DDS芯片,由它来完成基带信号的采样率转换、中频产生、调制和输出. 射频脉冲的相位和频率偏置的设置直接由级联NCO实现,而幅度设置则由步进衰减器和中频幅度调制共同实现. 该发射机的基本性能和功能满足常规1D/2D NMR实验需求.     

低场脉冲NMR中静磁场不均匀性的射频场补偿

在低场脉冲核磁共振实验中为了增大射频激励的带宽,通常采用的方法是提高射频激励磁场的场强. 针对共振区域中静磁场的不均匀性,本文提出了根据共振区域中的静磁场分布设计射频线圈以提高射频激励带宽,并用目标场方法实现了这一构想.      

Cyclic and Slice-Selective J Cross Polarization for Heteronuclear Editing and Localized NMR

This work has two objectives. First, heteronuclear editing was considered, with particular aim at proton-detected NMR spectroscopy of rare nuclei. The second aspect refers to localized heteronuclear NMR spectroscopy, implying the editing option. The common basis of both techniques is J cross polarization (or coherent rotating-frame polarization transfer). In the presence of magnetic field gradients this process is slice selective and is introduced as a principle for localized liquid-state NMR, particularly of heteronuclear spins. Heteronuclear editing of NMR lines is performed by the aid of a cyclic cross-polarization pathway. Both methods can be combined to a technique for localized proton-detected NMR spectroscopy of rare nuclei. Test experiments demonstrating the slice selectivity of the transfer process and the efficiency of the cyclic cross-polarization editing procedure are reported. Applications to localized homo- or heteronuclear spectroscopy and imaging are discussed.     

Recoupled Polarization-Transfer Methods for Solid-State H–C Heteronuclear Correlation in the Limit of Fast MAS

An in-depth account of the effects of homonuclear couplings and multiple heteronuclear couplings is given for a recently published technique for ¹H–¹³C dipolar correlation in solids under very fast MAS, where the heteronuclear dipolar coupling is recoupled by means of REDOR π-pulse trains. The method bears similarities to well-known solution-state NMR techniques, which form the framework of a heteronuclear multiple-quantum experiment. The so-called recoupled polarization-transfer (REPT) technique is versatile in that rotor-synchronized ¹H–¹³C shift correlation spectra can be recorded. In addition, weak heteronuclear dipolar coupling constants can be extracted by means of spinning sideband analysis in the indirect dimension of the experiment. These sidebands are generated by rotor encoding of the reconversion Hamiltonian. We present generalized variants of the initially described heteronuclear multiple-quantum correlation (HMQC) experiment, which are better suited for certain applications. Using these techniques, measurements on model compounds with ¹³C in natural abundance, as well as simulations, confirm the very weak effect of ¹H–¹H homonuclear couplings on the spectra recorded with spinning frequencies of 25–30 kHz. The effect of remote heteronuclear couplings on the spinning-sideband patterns of CH_n groups is discussed, and ¹³C spectral editing of rigid organic solids is shown to be practicable with these techniques.     

Double-Resonance Decoupling for Resolution Enhancement of P Solid-State MAS and Al → P MQHETCOR NMR

²⁷Al decoupling has been used to remove residual J-coupling interactions between³¹ P and ²⁷Al in microporous aluminophosphates AlPO₄-14 and AlPO₄-40. In combination with ¹H high-power decoupling, ²⁷Al adapted decoupling yields ³¹P spectra with optimal sensitivity and resolution. The importance of double-resonance decoupling is further demonstrated by incorporating this technique in the MQHETCOR sequence. Unambiguous assignment of all the AlPO₄-14 nuclear magnetic resonances is achieved by combining multiple-quantum evolution in the ²⁷Al dimension and double-resonance decoupling in the ³¹P acquisition domain.     

C Selective Polarization and Spin Diffusion in a Lipid Bilayer-Bound Polypeptide by Solid-State NMR

While ¹⁵N solid-state NMR has proven to be very advantageous for the development of structural biological methods, ¹³C spectroscopy has increased sensitivity and spectral dispersion. However, large natural abundance signals and homonuclear dipolar interactions pose significant problems. Here we have used a pair of ¹³C-labeled sites in a lipid-solubilized polypeptide to show the selective polarization can be used in combination with spin diffusion to achieve simplified spectra. Both unoriented and oriented samples have been used, with the latter providing a well-resolved homonuclear dipolar splitting.     

Fast acquisition of multi-dimensional spectra in solid-state NMR enabled by ultra-fast MAS

The advantages offered by ultra-fast (>60 kHz) magic angle spinning (MAS) rotation for the study of biological samples, notably containing paramagnetic centers are explored.It is shown that optimal conditions for performing solid-state ¹³C NMR under 60 kHz MAS are obtained with low-power CW ¹H decoupling, as well as after a low-power ¹H,¹³C cross-polarization step at a double-quantum matching condition. Acquisition with low-power decoupling highlights the existence of rotational decoupling sidebands. The sideband intensities and the existence of first and second rotary conditions are explained in the framework of the Floquet–van Vleck theory.As a result, optimal ¹³C spectra of the oxidized, paramagnetic form of human copper zinc superoxide dismutase (SOD) can be obtained employing rf-fields which do not exceed 40 kHz during the whole experiment. This enables the removal of unwanted heating which can lead to deterioration of the sample. Furthermore, combined with the short ¹H T₁s, this allows the repetition rate of the experiments to be shortened from 3 s to 500 ms, thus compensating for the sensitivity loss due to the smaller sample volume in a 1.3 mm rotor. The result is that 2D ¹³C–¹³C correlation could be acquired in about 24 h on less than 1 mg of SOD sample.     

低场核磁共振短死时间射频线圈与射频开关的设计

本文以核磁共振（NMR）射频线圈振铃信号产生原理为对象进行分析研究,提出了一种适用于低场环境下由环状间隙腔线圈与螺线管线圈构成的收发分离式短死时间射频线圈设计方案,采用优化调谐匹配网络提高发射效率;根据射频线圈方案需求设计了快速切换的射频开关及驱动．在此基础上依据仿真结果制作了短死时间射频线圈,并应用于自主研制的低场9.51 MHz便携式NMR谱仪系统,进行NMR实验,结果显示可将收发切换时间缩短至10 μs以内,验证了该设计方案的可行性．     

Temperature calibration for high-temperature MAS NMR to 913 K: 63Cu MAS NMR of CuBr and CuI, and 23Na MAS NMR of NaNbO3

The solid-state phase transitions of CuBr, CuI and NaNbO₃ can be readily observed using ⁶³Cu and ²³Na high-temperature magic-angle spinning nuclear magnetic resonance spectroscopy. Temperature has large, linear effects on the peak maximum of ⁶³Cu in each solid phase of CuBr and CuI, and there is large jump in shift across each phase transition. The ²³Na MAS NMR peak intensities and the line widths in NaNbO₃ also clearly show its high-temperature transition to the cubic phase. These data can be used to calibrate high-temperature MAS NMR probes up to 913 K, which is two hundred degrees higher than the commonly-used temperature calibration based on the chemical shift of ²⁰⁷Pb in Pb(NO₃)₂.     

基于半导体光放大器交叉偏振调制效应实现正、反相波长变换

利用琼斯矩阵,对基于半导体光放大器中的交叉偏振调制效应实现全光波长变换进行了理论分析.结果表明,利用这种机制可以实现同相与反相波长变换.在实验上同时实现了10Gb/s归零信号的正相与反相波长变换,输出消光比分别达到了6.9 dB和8.1 dB.     

Device Length Dependency of Cross Gain Modulation and Cross Phase Modulation in Semiconductor Optical Amplifier

The cross gain modulation, the cross phase modulation and their recovery time in the SOAs with the various lengths were experimentally investigated. It was found that these values strongly depended on the device length.     

Improved quantification of alite and belite in anhydrous Portland cements by Si MAS NMR: Effects of paramagnetic ions

The applicability, reliability, and repeatability of ²⁹Si MAS NMR for determination of the quantities of alite (Ca₃SiO₅) and belite (Ca₂SiO₄) in anhydrous Portland cement was investigated in detail for 11 commercial Portland cements and the results compared with phase quantifications based on powder X-ray diffraction combined with Rietveld analysis and with Taylor–Bogue calculations. The effects from paramagnetic ions (Fe³⁺) on the spinning sideband intensities, originating from dipolar couplings between ²⁹Si and the spins of the paramagnetic electrons, were considered and analyzed in spectra recorded at four magnetic fields (4.7–14.1 T) and this has led to an improved quantification of alite and belite from ²⁹Si MAS NMR spectra recorded at “high” spinning speeds of ν_R=12.0–13.0 kHz using 4 or 5 mm rotors. Furthermore, the impact of Fe³⁺ ions on the spin-lattice relaxation was studied by inversion-recovery experiments and it was found that the relaxation is overwhelmingly dominated by the Fe³⁺ ions incorporated as guest-ions in alite and belite rather than the Fe³⁺ sites present in the intimately mixed ferrite phase (Ca₂Al_xFe_2−xO₅).     

A general protocol for temperature calibration of MAS NMR probes at arbitrary spinning speeds

A protocol using (207)Pb NMR of solid lead nitrate was developed to determine the temperature of magic-angle spinning (MAS) NMR probes over a range of nominal set temperatures and spinning speeds. Using BioMAS and FastMAS probes with typical sample spinning rates of 8 and 35 kHz, respectively, empirical equations were devised to predict the respective sample temperatures. These procedures provide a straightforward recipe for temperature calibration of any MAS probe.     

A Fast Computation Technique for RF Performances in Broadband Millimeter Wave Helix TWTs

A computation technique for gain, efficiency and output power in broadband millimeter wave helix TWTs from their dimensions is described. The computed results by modeling are good agreement with experimental measurements. It is shown that the method is fast and useful accuracy. It can be used as engineering design for MMW TWTs.     

Fast Dissolution Dynamic Nuclear Polarization NMR of 13C-Enriched 89Y-DOTA Complex: Experimental and Theoretical Considerations

The yttrium complex of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetra(1′-¹³C-acetic acid) [¹³C]DOTA was synthesized. Fast dissolution dynamic nuclear polarization (DNP) nuclear magnetic resonance (NMR) studies revealed that the ⁸⁹Y, ¹³C, and ¹⁵N nuclei present in the complex could be co-polarized at the same optimum microwave irradiation frequency. The liquid-state spin–lattice relaxation time T ₁ of these nuclei were found to be reasonably long to preserve some or most of the DNP-enhanced polarization after dissolution. The hyperpolarized ¹³C and ⁸⁹Y NMR signals were optimized in different glassing mixtures. The overall results are discussed in light of the thermal mixing model of DNP.     

A Fast Three-Dimensional Protocol for Low-Field Laplace NMR in Porous Media

Three-dimensional (3-D) nuclear magnetic resonance (NMR) experiments reflect material structure, such as physical array of porous media and chemical compositions of complex fluids. However, a huge database is acquired in multi-dimensional NMR and obstructs the extraction of this information in a proper manner. In this paper, a new fast 3-D Laplace inversion procedure is introduced which contains a specially designed 3-D pulse sequence to acquire the data responding to interesting NMR properties simultaneously and efficient data-processing algorithm referring this pulse sequence. A 3-D NMR experiment in low-field on water-saturated synthetic porous sample is presented to demonstrate the validity of the designed 3-D inverse Laplace algorithm.