Department of Chemistry, University of Zurich, Winterthurerstrasse 190, 8057 Zurich (Switzerland) http://www.bioorganic-chemistry.com.
Pharmaceutical Biochemistry, University of Geneva (Switzerland).
Angew Chem Int Ed Engl. 2015 Jun 26;54(27):7911-4. doi: 10.1002/anie.201500250. Epub 2015 May 14.
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.
通过使用新型的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，可以实现病原体选择性标记。感染单纯疱疹病毒-1（HSV-1）的细胞，但未感染的细胞，在添加 dF-EdU 和荧光叠氮化物后会显示核染色。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 的活性部位，但空间位阻阻止 dF-EdU 与人类 TK 结合。这些结果提供了第一个例子，说明了在人类细胞中病原体-酶依赖性地掺入和标记生物正交官能团。
