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DEVELOPMENT OF A BIOSUPRAMOLECULAR NMR SYSTEM WITH A SUPERCONDUCTING MAGNET

具有超导磁体的生物超分子核磁共振系统的开发

基本信息

批准号：
03558029
负责人：
AKUTSU Hideo
金额：
$ 8.58万
依托单位：
YOKOHAMA NATIONAL UNIVERSITY (Y. N. U.)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1992
项目状态：
已结题

项目摘要

A multinuclear observation system was built into our AM-400 NMR spectrometer (Bruker). Then, a solid-state probe head for deuterium NMR was developed. The quadrupole echo pulse sequence was used for the measurements. At first, a solenoid coil was built in a probe head, originally used for solution NMR. The 90ﾟ pulse was 12mus. The circuits of the probe head and matching box were simplified. After the modification, the 90ﾟ pulse became to be about 4mus. Thus, we have established the solid-state NMR system, which can be used for the start point of this project. phospholipids specifically deuterated in the glycerol backbone were obtained from E. coli cells cultured in the presence of perdeuterated glycerol.Since biological samples contain water, the sample tube should be held vertically. Although the power is very efficient in the solenoid coil, the sample tube is kept in horizontal. So we decided to change the coil design. A saddle type coil was attached on the surface of a glass tube with 5 mmphi, which was put in the vertical direction. The 8-turn coil fits to 0.94 cm^3 sample volume. The 90ﾟ pulse of this coil was 7.8mus. In order to get more power, a high power amplifier was connected. However, arching took place in the probe head on the use of this amplifier. Since the bore size of our magnet is narrow, the probe is not enough to avoid the arching. The probe was examined without the amplifier. By using 60 mg deuterated phospholipids, a ^2H-NMR spectrum in good quality was obtained. The spin-lattice relaxation time was measured using the inversion-recovery method. Whole region of the power pattern spectrum was inverted by the 180ﾟ pulse. These results show that this coil can be used for the investigation of the phospholipid bilayers. Thus, we have succeeded in establishing a solid-state NMR system suitable for biomembrane studies.

我们的 AM-400 NMR 波谱仪 (Bruker) 内置了多核观测系统。然后，开发了用于氘核磁共振的固态探头。四极回波脉冲序列用于测量。最初，螺线管线圈内置于探头中，最初用于溶液核磁共振。 90ﾟ脉冲为12mus。简化了探头和匹配盒的电路。修改后，90ﾟ脉冲变为4mus左右。因此，我们建立了固态核磁共振系统，可以作为该项目的起点。甘油主链中专门氘化的磷脂是从在全氘化甘油存在下培养的大肠杆菌细胞中获得的。由于生物样品含有水，因此样品管应垂直放置。尽管电磁线圈的功率非常有效，但样品管仍保持水平。因此我们决定改变线圈设计。将鞍型线圈安装在5mmphi玻璃管的表面上，将其垂直放置。 8 匝线圈适合 0.94 cm^3 样品体积。该线圈的90ﾟ脉冲为7.8mus。为了获得更大的功率，连接了高功率放大器。然而，使用该放大器时，探头头部发生了电弧现象。由于我们的磁铁孔径很窄，探头不足以避免拱形。在没有放大器的情况下检查探头。通过使用 60 mg 氘化磷脂，获得了质量良好的 2H-NMR 谱。使用反转恢复方法测量自旋晶格弛豫时间。功率模式频谱的整个区域被 180ﾟ脉冲反转。这些结果表明该线圈可用于磷脂双层的研究。因此，我们成功建立了适合生物膜研究的固态核磁共振系统。