TRAINING IN USE OF DMX ELECTRONICS & NMR

~1H核磁共振溶液结构的测定过程 荷兰P.Hollandica塑料花青素(PC)的制备将包括若干步骤， 从获取二维1H-1H相关核磁共振开始 荷兰P.Hollandica PC在H2O中的DQF-COSY、TOCSY和NOESY光谱 不同的温度，通过使用标准脉冲序列。保龄球 格林州立大学在现场安装了一台400 MHz的瓦里安核磁共振光谱仪， 并将在适当的时候从更高的地方获得额外的光谱 位于别处的现场仪器。水信号抑制将 通常通过预饱和或脉冲场梯度来实现。这个 上述光谱也将在D2O中获取，以确定 慢速和快速交换酰胺质子和鉴定β-质子 否则就会被水照射浸透。序列特异性共振 作业将通过标准程序获得。如果需要， 使用1H3D将解决信号重叠造成的模糊性 TOCSY-NOESY谱。距离和二面角约束 将从核Overhauser效应和标量耦合中评估 常量。获取的约束将用于生成 程序X-PLOR中的结构。结构计算和结构计算 NOESY信号分配将迭代执行。接下来， 产生的结构将通过咒语能量最小化或 能量最小化与约束分子动力学的结合 方法：研究方法。荷兰P.Hollandica和其他PC结构的分析将 使用广达程序(Biosym/MSI，圣地亚哥)进行。核磁共振 数据分析、约束条件的生成和比较 实验NOESY谱与反演谱的结合 生成的结构将使用Felix执行 (Biosym/MSI，圣地亚哥)。

The process of the determination of the 1H NMR solution structure of P. hollandica plastocyanin (PC) will include several steps, beginning with the acquisition of two-dimnsional 1H-1H correlated NMR spectra (DQF-COSY, TOCSY, and NOESY) of P. hollandica PC in H2O at different temperatures, by using standard pulse sequences. Bowling Green State University has a 400 MHz Varian NMR spectrometer on site, and additional spectra will be acquired when appropriate from higher field instruments located elsewhere. Water signal suppression will typically be achieved by presaturation or pulse-field gradient. The above mentioned spectra will also be acquired in D2O to identify the slow and fast exchanging amide protons and to identify the ?-protons otherwise saturated by water irradiation. Sequence-specific resonance assignments will be obtained by standard procedures. If required, the ambiguities caused by signal overlaps will be solved by using 1H 3D TOCSY-NOESY spectrum. The distance and dihedral angle constraints will be eveluated from nuclear Overhauser effects and scalar coupling constants. The obtained constraints will be used to generate structures in the program X-PLOR. The structure calculations and the NOESY signal assignments will be performed iteratively. Next, the resulting structures will be refined by CHARM energy minimization or a combination of energy minimization and restrained molecular dynamics methods. The analysis of P. hollandica and other PC structures will be performed using the program QUANTA (Biosym/MSI, San Diego). NMR data analysis, restraints generation and comparison of the experimental NOESY spectra with the back-calculated spectra together with the structure generated will be carried out using FELIX (Biosym/MSI, San Diego).