Leibniz Institute of Molecular Pharmacology (FMP Berlin), Robert Roessle Strasse 10, 13125 Berlin, Germany.
J Am Chem Soc. 2012 May 9;134(18):7616-9. doi: 10.1021/ja301895f. Epub 2012 Apr 27.
Methylation and acetylation of protein lysine residues constitute abundant post-translational modifications (PTMs) that regulate a plethora of biological processes. In eukaryotic proteins, lysines are often mono-, di-, or trimethylated, which may signal different biological outcomes. Deconvoluting these different PTM types and PTM states is not easily accomplished with existing analytical tools. Here, we demonstrate the unique ability of NMR spectroscopy to discriminate between lysine acetylation and mono-, di-, or trimethylation in a site-specific and quantitative manner. This enables mapping and monitoring of lysine acetylation and methylation reactions in a nondisruptive and continuous fashion. Time-resolved NMR measurements of different methylation events in complex environments including cell extracts contribute to our understanding of how these PTMs are established in vitro and in vivo.
蛋白质赖氨酸残基的甲基化和乙酰化构成了丰富的翻译后修饰（PTMs），调节着大量的生物学过程。在真核蛋白质中，赖氨酸常被单、二或三甲基化，这可能标志着不同的生物学结果。利用现有的分析工具，很难将这些不同的 PTM 类型和 PTM 状态区分开来。在这里，我们展示了 NMR 光谱学以特定和定量的方式区分赖氨酸乙酰化与单、二或三甲基化的独特能力。这使得在非破坏性和连续的方式下对赖氨酸乙酰化和甲基化反应进行作图和监测成为可能。在包括细胞提取物在内的复杂环境中对不同甲基化事件的时间分辨 NMR 测量有助于我们理解这些 PTM 是如何在体外和体内建立的。
