13C LABELED GLYCEROL

SIR提供[13C，1，3‘-H4]甘油；14g均一的13C 富集物已被广泛用于共振指认研究。 蛋白质，以及最近的核酸。以及各种 好处，这种标签模式有几个限制， 源于存在一个键13C-13C偶联。除了……之外 13C维的退化解只能部分地 通过恒定时间技术缓解，未检测到13C 开展了松弛实验，证明了松弛实验的有效性。 由于13C-13C标量相互作用而产生的人工产物。交替的 我们开发的用于大肠杆菌表达的碳富集法 系统允许松弛实验以及各种13C 编辑作业实验。这种方法包括培养 以[2-13C]-或[1，3-13C21甘油]作为碳源的生物体。 此外，我们还证明了通过将随机分数 通过选择性13C浓缩，人们可以分析检测到的黄体生成素 亚甲基碳的松弛，这是到目前为止还没有 可行。结果是通过使用13C的两个相反的图案 应该可以确定运动参数。 几乎所有的蛋白质或核酸分子的位置。在……里面 此外，特别是在核酸的情况下，我们预计 这种选择性标签的方法将提供实质性的 通过均匀的~(13)C浓缩补充结构研究。

The SIR provided [13C, 1,3-'H4 ]Glycerol; 14g Uniform 13C enrichment has become widely used for resonance assignment studies of proteins and, more recently, nucleic acids. Along with the various benefits, there are several limitations to this labeling pattern which stem from the presence of one bond 13C-13C couplings. In addition to the degraded reslution in the 13C dimension which can only partially be alleviated by constant time techniques, no inderct detected 13C relaxation experiments have been developed which cercumvent the artifacts due to the 13C-13C scalar interactions. The alternating carbon enrichment procedure we have developed for E. coli expression systems allows for both relaxation experiments as well as various 13C editing assignment experiments. This method involves culturing the organisms using [2-13C]- or [1,3-13C21glycerol as a carbon source. Furthermore, we have demonstrated that by combining random fractional deuteration with selective 13C enrichment, one can analyze lH detected relaxation of methylene carbons which has heretofore not been feasible. As a result by using the two opposite patterns of 13C enrichment it should be possible to deterine the motional parameters of nearly all sites of a protein or nucleic acid molecule. In addition, particularly in the case of nucleic acids, we anticipate that this approach to selective labeling will offer a substantial complement to structural studies via uniform 13C enrichment.