C-13 MAGIC-ANGLE SAMPLE-SPINNING NUCLEAR-MAGNETIC-RESONANCE STUDIES OF HUMAN MYELIN, AND MODEL MEMBRANE SYSTEMS
C-13 MAGIC-ANGLE SAMPLE-SPINNING NUCLEAR-MAGNETIC-RESONANCE STUDIES OF HUMAN MYELIN, AND MODEL MEMBRANE SYSTEMS
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DOI：
10.1002/mrm.1910290204
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发表时间：
1993-02-01
影响因子：
3.3
通讯作者：
OLDFIELD, E
中科院分区：
文献类型：
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作者：
HUSTED, C;MONTEZ, B;OLDFIELD, E
我们已经获得了高场（11.7特斯拉），高分辨率的碳-13固态“魔角”样品旋转核磁共振（NMR）光谱的各种磷脂，鞘脂，髓鞘和白色物质样品，解决和分配超过40共振的光谱中的人类和牛髓鞘。NMR结果表明，没有大的光谱变化，由于样品制备，样品冷冻，或大脑的位置，也没有髓鞘结构的变化，可通过光学显微镜，电子显微镜，薄层色谱法，或十二烷基硫酸钠-聚丙烯酰胺凝胶电泳检测，归因于有时冗长的NMR数据采集过程。人类髓鞘和白色物质的化学位移分配的基础上的C-13“魔角”样品旋转（MAS）NMR光谱的个别模型脂质，以及对光谱的脂质混合物。在所有髓磷脂样品中，基本上没有可归因于膜蛋白的特征，除了一个小的特征，由于Czeta的精氨酸残基，主要是在髓磷脂碱性蛋白。模型脂质和完整髓鞘光谱之间的一般相似性表明蛋白质对脂质流动性没有重大影响。我们还研究了人类髓磷脂样品作为发育年龄（4，15，48个月和成人）的函数，我们的结果显示，尽管髓磷脂和模型化合物的计算机线形模拟确定，脂质烃链不饱和度随着年龄的增长而显著降低，但总体脂质组成只有很小的变化。婴儿脑白质病髓鞘谱显示半乳糖苷脂明显减少。总的来说，能够解决和分配超过40个共振的C-13 MAS NMR光谱的髓鞘，并检测变化的功能的发展和疾病，应该提供一个有用的起点，进一步更详细的研究髓鞘膜分子运动和功能。
We have obtained high-field (11.7 Tesla), high-resolution carbon-13 solid-state ''magic-angle'' sample-spinning nuclear magnetic resonance (NMR) spectra of a variety of phospholipids, sphingolipids, myelin and white matter samples, resolving and assigning over 40 resonances in the spectra of human and bovine myelin. The NMR results indicated no large spectral changes due to sample preparation, sample freezing, or brain location, and also no changes in myelin structure detectable via light microscopy, electron microscopy, thin layer chromatography, or sodium dodecyl sulfate-polyacrylamide gel electrophoresis, attributable to the sometimes lengthy NMR data acquisition process. Human myelin and white matter chemical shift assignments were made based on C-13 ''magic angle'' sample spinning (MAS) NMR spectra of individual model lipids, as well as on spectra of lipid mixtures. In all myelin samples there were essentially no features attributable to membrane proteins, with the exception of one small feature due to Czeta of Arg residues, primarily in the myelin basic proteins. The general similarity between the model lipid and intact myelin spectra suggested no major effects of protein on lipid mobility. We have also investigated human myelin samples as a function of developmental age (4,15, 48 months and adult), and our results showed only small changes in overall lipid composition, although there were significant decreases in lipid hydrocarbon chain unsaturation with age, as determined by computer line-shape simulations of myelin and model compounds. The spectrum of an infant leukoencephalopathy myelin showed marked decreases in galactocerebrosides. Overall, the ability to resolve and assign over 40 resonances in the C-13 MAS NMR spectra of myelin, and to detect changes as a function of development and disease, should provide a useful starting point for further more detailed studies of myelin membrane molecular motions, and function.