A simple biosynthetic method for stereospecific resonance assignment of prochiral methyl groups in proteins
A simple biosynthetic method for stereospecific resonance assignment of prochiral methyl groups in proteins
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DOI：
10.1007/s10858-010-9463-3
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发表时间：
2011-02-01
影响因子：
2.7
通讯作者：
Gans, Pierre
中科院分区：
文献类型：
--
作者：
Plevin, Michael J.;Hamelin, Olivier;Gans, Pierre
本文提出了一种新的立体专一性地确定蛋白质中前手性甲基的方法。该方法是用U-[C-13]葡萄糖和亚饱和量的2-[C-13]甲基乙酰乳酸制备蛋白质样品。由此产生的不均匀的标记图案允许proR和proS甲基基团可以很容易地区分它们的不同阶段中的恒定时间的二维H-1-C-13相关光谱。使用与主要均匀标记的样品相同的介质组成方便地制备蛋白质样品，并且比分步标记方法含有更高水平的同位素富集。因此，这种新的策略代表了一个经济上有吸引力的，强大的替代获得同位素编码的立体特异性NMR分配的前手性甲基。
A new method for stereospecific assignment of prochiral methyl groups in proteins is presented in which protein samples are produced using U-[C-13]glucose and subsaturating amounts of 2-[C-13]methyl-acetolactate. The resulting non-uniform labeling pattern allows proR and proS methyl groups to be easily distinguished by their different phases in a constant-time two-dimensional H-1-C-13 correlation spectra. Protein samples are conveniently prepared using the same media composition as the main uniformly-labeled sample and contain higher levels of isotope-enrichment than fractional labeling approaches. This new strategy thus represents an economically-attractive, robust alternative for obtaining isotopically-encoded stereospecific NMR assignments of prochiral methyl groups.