通过核磁共振氢谱和中子散射法估算全细胞和细胞蛋白质中的氘代水平。
Estimation of deuteration levels in whole cells and cellular proteins by 1H n.m.r. spectroscopy and neutron scattering.
作者信息
Perkins S J
出版信息
Biochem J. 1981 Oct 1;199(1):163-70. doi: 10.1042/bj1990163.
Abstract
Methods using conventional Fourier transform 1H n.m.r. spectroscopy at 250 MHz for the determination of the overall deuteration levels of cells cultured in media containing 2H2O or deuterated carbon sources are described. These were developed for Escherichia coli as a model, and extended to Neurospora crassa hyphae and mouse myeloma cells P815. The results were investigated by 1H n.m.r. and neutron scattering measurements on deuterated proteins that were obtained from E. coli. It is concluded that 1H n.m.r. is able to observe the soluble proteins of E. coli in certain cases, that deuteration levels can be determined by 1H n.m.r. for small quantities of proteins in their native state, and that glycerol is a more efficient carbon source than glucose for the deuteration of E. coli proteins.
摘要
描述了使用250兆赫常规傅里叶变换1H核磁共振光谱法来测定在含有2H2O或氘代碳源的培养基中培养的细胞的整体氘化水平的方法。这些方法是以大肠杆菌为模型开发的，并扩展到粗糙脉孢菌菌丝和小鼠骨髓瘤细胞P815。通过对从大肠杆菌获得的氘代蛋白质进行1H核磁共振和中子散射测量来研究结果。得出的结论是，1H核磁共振在某些情况下能够观察到大肠杆菌的可溶性蛋白质，对于少量天然状态的蛋白质，1H核磁共振能够测定其氘化水平，并且甘油是比葡萄糖更有效的用于大肠杆菌蛋白质氘化的碳源。
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