55520-67-7Relevant articles and documents
Alkene-tetrazine ligation for imaging cellular DNA
Rieder, Ulrike,Luedtke, Nathan W.
, p. 9168 - 9172,5 (2014)
5-Vinyl-2′-deoxyuridine (VdU) is the first reported metabolic probe for cellular DNA synthesis that can be visualized by using an inverse electron demand Diels-Alder reaction with a fluorescent tetrazine. VdU is incorporated by endogenous enzymes into the genomes of replicating cells, where it exhibits reduced genotoxicity compared to 5-ethynyl-2′-deoxyuridine (EdU). The VdU-tetrazine ligation reaction is rapid (k≈0.02 M-1 s -1) and chemically orthogonal to the alkyne-azide "click" reaction of EdU-modified DNA. Alkene-tetrazine ligation reactions provide the first alternative to azide-alkyne click reactions for the bioorthogonal chemical labeling of nucleic acids in cells and facilitate time-resolved, multicolor labeling of DNA synthesis. Diels-Alder on DNA: 5-Vinyl-2′-deoxyuridine (VdU) is metabolically incorporated into cellular DNA where it can be visualized by using inverse electron demand Diels-Alder reactions with fluorescent tetrazines. VdU-tetrazine ligation reactions are rapid (k≈0.02 M-1 s-1) and chemically orthogonal to alkyne-azide "click" reactions, thereby enabling time-resolved, multicolor labeling of DNA synthesis in individual cells.
One-pot approach to functional nucleosides possessing a fluorescent group using nucleobase-exchange reaction by thymidine phosphorylase
Hatano, Akihiko,Kurosu, Masayuki,Yonaha, Susumu,Okada, Munehiro,Uehara, Sanae
, p. 6900 - 6905 (2013/10/08)
Herein, we describe β-selective coupling between a modified uracil and a deoxyribose to produce functionalized nucleosides catalyzed by thymidine phosphorylase derived from Escherichia coli. This enzyme mediates nucleobase-exchange reactions to convert unnatural nucleosides possessing a large functional group such as a fluorescent molecule, coumarin or pyrene, linked via an alkyl chain at the C5 position of uracil. 5-(Coumarin-7-oxyhex-5- yn)uracil (C4U) displayed 57.2% conversion at 40% DMSO concentration in 1.0 mM phosphate buffer pH 6.8 to transfer thymidine to an unnatural nucleoside with C4U as the base. In the case of using 5-(pyren-1-methyloxyhex-5-yn)uracil (P4U) as the substrate, TP also could catalyse the reaction to generate a product with a very large functional group at 50% DMSO concentration (21.6% conversion). We carried out docking simulations using MF myPrest for the modified uracil bound to the active site of TP. The uracil moiety of the substrate binds to the active site of TP, with the fluorescent moiety linked to the C5 position of the nucleobase located outside the surface of the enzyme. As a consequence, the bulky fluorescent moiety binding to uracil has little influence on the coupling reaction.
Detection of methylcytosine by DNA photoligation via hydrophobic interaction of the alkyl group
Ogino, Masayuki,Taya, Yuuta,Fujimoto, Kenzo
supporting information; experimental part, p. 3163 - 3167 (2011/02/25)
We report the nonenzymatic detection of 5-methylcytosine by using template-directed photoligation through 5-vinyl-2′-deoxyuridine derivatives with high selectivity. In this paper, we propose a new detection method by using hydrophobic interaction. The photoligation yield of the 5-methylcytosine case was approximately 5.6-fold higher than that in the case of cytosine.
