Preparation of Recombinant Peptides with Site- and Degree-Specific Lysine 13C-Methylation
Preparation of Recombinant Peptides with Site- and Degree-Specific Lysine 13C-Methylation
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DOI：
10.1021/bi900348z
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发表时间：
2009-05-12
期刊：
影响因子：
2.9
通讯作者：
Mer, Georges
中科院分区：
文献类型：
--
作者：
Cui, Gaofeng;Botuyan, Maria Victoria;Mer, Georges
赖氨酸甲基化是影响蛋白质功能的重要翻译后修饰;例如。p53肿瘤抑制蛋白的转录活性。为了便于结构表征的复合物涉及蛋白质和甲基化的目标，通过核磁共振光谱，我们设计了一种简单的方法，用于制备重组N-15/C-13-富集肽与C-13-甲基标记的甲基化赖氨酸类似物。该方法，它依赖于C-13-富集的烷化剂的合成，被施加到生产的15个残基的p53肽在赖氨酸类似物370不同甲基化。这些肽用于探测单、二和三甲基化p53与三种不同蛋白质的甲基化状态依赖性相互作用。
Lysine methylation is an important post-translational modification that affects protein function; for example.. the transcriptional activity of the p53 tumor suppressor protein. To facilitate structural characterization of complexes involving proteins and methylated targets by nuclear magnetic resonance spectroscopy, we devised a simple method for preparing recombinant N-15/C-13-enriched peptides with a C-13-methyl-labeled methylated lysine analogue. The method, which relies on the synthesis of C-13-enriched alkylating agents, was applied to the production of 15-residue p53 peptides variously methylated at lysine analogue 370. The peptides were used to probe the methylation state-dependent interactions of mono, di, and trimethylated p53 with three different proteins.