Selective 1H-13C NMR spectroscopy of methyl groups in residually protonated samples of large proteins
Selective 1H-13C NMR spectroscopy of methyl groups in residually protonated samples of large proteins
复制标题
DOI：
10.1007/s10858-009-9393-0
复制
发表时间：
2010-02-01
影响因子：
2.7
通讯作者：
Tugarinov, Vitali
中科院分区：
文献类型：
--
作者：
Guo, Chenyun;Tugarinov, Vitali
所有含甲基氨基酸的甲基（CHD 2）-C-13同位素异构体可以在从基于[U-C-13，H-1]-葡萄糖/D2 O的细菌培养基获得的大蛋白质的残留质子化样品中观察到，其灵敏度足以用于许多NMR应用。使用简单的“前后”NMR方法可以选择性地检测甲基的一些子集（Ala（beta）、Thr（gamma 2））。这种选择性甲基检测的“前后”NMR实验允许在82-kDa酶苹果酸合酶G的残留质子化的[U-C-13，H-1]-葡萄糖/D2 O衍生样品中完全指定苏氨酸γ 2甲基。[U-C-13，H-1]-葡萄糖/D2 O衍生蛋白质样品相对便宜，通常在高分子量系统的任何NMR研究的早期阶段就可获得。
Methyl (CHD2)-C-13 isotopomers of all methyl-containing amino-acids can be observed in residually protonated samples of large proteins obtained from [U-C-13,H-1]-glucose/D2O-based bacterial media, with sensitivity sufficient for a number of NMR applications. Selective detection of some subsets of methyl groups (Ala(beta), Thr(gamma 2)) is possible using simple 'out-and-back' NMR methodology. Such selective methyl-detected 'out-and-back' NMR experiments allow complete assignments of threonine gamma 2 methyls in residually protonated, [U-C-13,H-1]-glucose/D2O-derived samples of an 82-kDa enzyme Malate Synthase G. [U-C-13,H-1]-glucose/D2O-derived protein samples are relatively inexpensive and are usually available at very early stages of any NMR study of high-molecular-weight systems.