Lee A L, Urbauer J L, Wand A J
Department of Biological Sciences, State University of New York at Buffalo 14260-3000, USA.
J Biomol NMR. 1997 Jun;9(4):437-40. doi: 10.1023/a:1018311013338.
Selective incorporation of 13C into the methyl groups of protein side chains is described as a means for simplifying the measurement and interpretation of 13C relaxation parameters. High incorporation (> 90%) is accomplished by using pyruvate (3-13C, 99%) as the sole carbon source in the growth media for protein overexpression in E. coli. This improved labeling scheme increases the sensitivity of the relaxation experiments by approximately fivefold when compared to randomly fractionally 13C-labeled protein, allowing high-quality measurements on relatively dilute (< 1 mM) protein samples at a relatively low cost.
将¹³C选择性掺入蛋白质侧链的甲基中，被描述为一种简化¹³C弛豫参数测量和解释的方法。通过使用丙酮酸（3-¹³C，99%）作为大肠杆菌中蛋白质过表达生长培养基中的唯一碳源，可实现高掺入率（>90%）。与随机部分¹³C标记的蛋白质相比，这种改进的标记方案将弛豫实验的灵敏度提高了约五倍，从而能够以相对较低的成本对相对稀释（<1 mM）的蛋白质样品进行高质量测量。
