Improving the fluorescent probe acridonylalanine through a combination of theory and experiment
Improving the fluorescent probe acridonylalanine through a combination of theory and experiment
复制标题
理论与实验相结合改进荧光探针吖啶丙氨酸
DOI：
10.1002/poc.3813
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发表时间：
2018
影响因子：
1.8
通讯作者：
Kohli, Rahul M.
中科院分区：
文献类型：
--
作者：
Sungwienwong, Itthipol;Ferrie, John J.;Jun, Joomyung V.;Liu, Chunxiao;Barrett, Taylor M.;Hostetler, Zachary M.;Ieda, Naoya;Hendricks, Amara;Muthusamy, Anand K.;Kohli, Rahul M.
吖啶丙氨酸 (Acd) 是一种有用的荧光团，可用于通过荧光光谱研究蛋白质，但它可以通过更长的波长或更亮来改进。在这里，我们报告了 Acd 核心衍生物的合成及其光物理表征。我们还对 Acd 衍生物的吸收和发射光谱进行了从头计算，这与实验测量结果非常吻合。氨基酸氨基吖啶基丙氨酸（Aad）以适合遗传掺入和肽合成的形式合成。我们在肽裂解测定中表明，Aad 是优于 Acd 的 Förster 共振能量转移受体，并且 Aad 可以被氨酰 tRNA 合成酶激活以进行基因掺入。总之，这些结果表明我们可以使用计算来设计增强的 Acd 衍生物，这些衍生物可用于肽和蛋白质。
Acridonylalanine (Acd) is a useful fluorophore for studying proteins by fluorescence spectroscopy, but it can potentially be improved by being made longer wavelength or brighter. Here, we report the synthesis of Acd core derivatives and their photophysical characterization. We also performed ab initio calculations of the absorption and emission spectra of Acd derivatives, which agree well with experimental measurements. The amino acid aminoacridonylalanine (Aad) was synthesized in forms appropriate for genetic incorporation and peptide synthesis. We show that Aad is a superior Förster resonance energy transfer acceptor to Acd in a peptide cleavage assay and that Aad can be activated by an aminoacyl tRNA synthetase for genetic incorporation. Together, these results show that we can use computation to design enhanced Acd derivatives, which can be used in peptides and proteins.