Rapid Structural Analysis of a Synthetic Non-canonical Amino Acid by Microcrystal Electron Diffraction.
Rapid Structural Analysis of a Synthetic Non-canonical Amino Acid by Microcrystal Electron Diffraction.
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DOI：
10.3389/fmolb.2020.609999
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发表时间：
2020
影响因子：
5
通讯作者：
Mills JH
中科院分区：
文献类型：
--
作者：
Gleason PR;Nannenga BL;Mills JH
小分子的结构表征是有机合成的重要组成部分。在这项工作中，我们应用微晶电子衍射（MicroED）分析了酶促反应产物的结构，该反应旨在产生非天然氨基酸2,4-二羟基苯丙氨酸（24DHF）。用核磁共振波谱（NMR）和质谱（MS）对我们分离的产物进行表征表明，已经形成了24DHF的异构体。然后用分离产物中的微晶以0.62 Å分辨率测定其结构，确定其为2-氨基-2-（2,4-二羟基苯基）丙酸（24DHPA）。此外，MicroED结构模型表明，不对称单元中存在两种对映体形式的24DHPA。值得注意的是，整个结构测定过程包括设置、数据收集和细化在1小时内完成。MicroED数据不仅支持了之前使用NMR和MS获得的结果，而且还立即提供了产品中存在的立体异构体的信息，这是单独使用NMR和MS难以实现的。因此，我们的研究结果表明，MicroED方法可以在类似于许多常用分析方法的时间尺度上提供有用的结构信息，并且可以在未来的研究中添加到现有的小分子结构测定工具中。
Structural characterization of small molecules is a crucial component of organic synthesis. In this work, we applied microcrystal electron diffraction (MicroED) to analyze the structure of the product of an enzymatic reaction that was intended to produce the unnatural amino acid 2,4-dihydroxyphenylalanine (24DHF). Characterization of our isolated product with nuclear magnetic resonance spectroscopy (NMR) and mass spectrometry (MS) suggested that an isomer of 24DHF had been formed. Microcrystals present in the isolated product were then used to determine its structure to 0.62 Å resolution, which confirmed its identity as 2-amino-2-(2,4-dihydroxyphenyl)propanoic acid (24DHPA). Moreover, the MicroED structural model indicated that both enantiomeric forms of 24DHPA were present in the asymmetric unit. Notably, the entire structure determination process including setup, data collection, and refinement was completed in ~1 h. The MicroED data not only bolstered previous results obtained using NMR and MS but also immediately provided information about the stereoisomers present in the product, which is difficult to achieve using NMR and MS alone. Our results therefore demonstrate that MicroED methods can provide useful structural information on timescales that are similar to many commonly used analytical methods and can be added to the existing suite of small molecule structure determination tools in future studies.
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影响因子：
16.8
作者：
Gallagher-Jones M;Glynn C;Boyer DR;Martynowycz MW;Hernandez E;Miao J;Zee CT;Novikova IV;Goldschmidt L;McFarlane HT;Helguera GF;Evans JE;Sawaya MR;Cascio D;Eisenberg DS;Gonen T;Rodriguez JA
通讯作者：
Rodriguez JA
低复杂性蛋白段的原子结构揭示了组装网络的扭结β薄片。
DOI：
10.1126/science.aan6398
发表时间：
2018-02-09
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Hughes MP;Sawaya MR;Boyer DR;Goldschmidt L;Rodriguez JA;Cascio D;Chong L;Gonen T;Eisenberg DS
通讯作者：
Eisenberg DS
影响因子：
48
作者：
Nannenga, Brent L.;Shi, Dan;Leslie, Andrew G. W.;Gonen, Tamir
通讯作者：
Gonen, Tamir
影响因子：
6.1
作者：
Hübschle CB;Sheldrick GM;Dittrich B
通讯作者：
Dittrich B
影响因子：
15
作者：
Dick, Markus;Sarai, Nicholas S.;Arnold, Frances H.
通讯作者：
Arnold, Frances H.