A Bioorthogonal Chemical Reporter of Viral Infection.
A Bioorthogonal Chemical Reporter of Viral Infection.
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DOI：
10.1002/ange.201500250
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发表时间：
2015-06-26
期刊：
影响因子：
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通讯作者：
Luedtke, Nathan W
中科院分区：
文献类型：
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作者：
Neef, Anne B;Pernot, Lucile;Luedtke, Nathan W
通过使用新型吉西他滨代谢物类似物2 '-脱氧-2'，2 '-二氟-5-乙炔基尿苷（dF-EdU）和点击化学实现病原体选择性标记。在加入dF-EdU和荧光叠氮化物后，感染单纯疱疹病毒-1（HSV-1）的细胞（而非未感染的细胞）表现出核染色。将dF-EdU掺入DNA取决于其被疱疹病毒胸苷激酶（TK）磷酸化。晶体学分析揭示了dF-EdU如何很好地适应HSV-1 TK的活性位点，但空间冲突阻止dF-EdU结合人TK。这些结果提供了第一个例子，病原体酶依赖的掺入和标记的生物正交功能团在人类细胞。
Pathogen-selective labeling was achieved by using the novel gemcitabine metabolite analogue 2'-deoxy-2',2'-difluoro-5-ethynyluridine (dF-EdU) and click chemistry. Cells infected with Herpes Simplex Virus-1 (HSV-1), but not uninfected cells, exhibit nuclear staining upon the addition of dF-EdU and a fluorescent azide. The incorporation of the dF-EdU into DNA depends on its phosphorylation by a herpes virus thymidine kinase (TK). Crystallographic analyses revealed how dF-EdU is well accommodated in the active site of HSV-1 TK, but steric clashes prevent dF-EdU from binding human TK. These results provide the first example of pathogen-enzyme-dependent incorporation and labeling of bioorthogonal functional groups in human cells.