Synthesis of a 13C-methylene-labeled isoleucine precursor as a useful tool for studying protein side-chain interactions and dynamics

IF 16.4 1区 化学 Q1 CHEMISTRY, MULTIDISCIPLINARY
Theresa Höfurthner, Giorgia Toscano, Georg Kontaxis, Andreas Beier, Moriz Mayer, Leonhard Geist, Darryl B. McConnell, Harald Weinstabl, Roman Lichtenecker, Robert Konrat
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Abstract

In this study, we present the synthesis and incorporation of a metabolic isoleucine precursor compound for selective methylene labeling. The utility of this novel α-ketoacid isotopologue is shown by incorporation into the protein Brd4-BD1, which regulates gene expression by binding to acetylated histones. High quality single quantum 13C−1 H-HSQC were obtained, as well as triple quantum HTQC spectra, which are superior in terms of significantly increased 13C-T2 times. Additionally, large chemical shift perturbations upon ligand binding were observed. Our study thus proves the great sensitivity of this precursor as a reporter for side-chain dynamic studies and for investigations of CH-π interactions in protein-ligand complexes.

Abstract Image

合成13C-亚甲基标记的异亮氨酸前体,作为研究蛋白质侧链相互作用和动力学的有用工具。
在本研究中,我们提出了一种用于选择性亚甲基标记的代谢异亮氨酸前体化合物的合成和掺入。这种新型α-酮酸异拓扑结构的实用性通过掺入蛋白质Brd4-BD1来显示,该蛋白质通过与乙酰化组蛋白结合来调节基因表达。获得了高质量的单量子13C-1 H-HSQC以及三量子HTQC光谱,其在显著增加13C-T2倍方面是优越的。此外,观察到配体结合的大的化学位移扰动。因此,我们的研究证明了这种前体作为侧链动力学研究和蛋白质-配体复合物中CH-π相互作用研究的报告物的高度敏感性。
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来源期刊
Accounts of Chemical Research
Accounts of Chemical Research 化学-化学综合
CiteScore
31.40
自引率
1.10%
发文量
312
审稿时长
2 months
期刊介绍: Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.
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