One-pot approach to functional nucleosides possessing a fluorescent group using nucleobase-exchange reaction by thymidine phosphorylase

Akihiko Hatano, Masayuki Kurosu, Susumu Yonaha, Munehiro Okada, Sanae Uehara

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)6900-6905
Number of pages6
JournalOrganic and Biomolecular Chemistry
Volume11
Issue number40
DOIs
Publication statusPublished - 2013 Oct 28

Fingerprint

Thymidine Phosphorylase
thymidine
nucleosides
uracil
Uracil
Nucleosides
Dimethyl Sulfoxide
Functional groups
enzymes
Deoxyribose
pyrenes
Substrates
Enzymes
Escherichia
Thymidine
Escherichia coli
phosphates
Buffers
buffers
Phosphates

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Biochemistry

Cite this

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.

In: Organic and Biomolecular Chemistry, Vol. 11, No. 40, 28.10.2013, p. 6900-6905.

Research output: Contribution to journalArticle

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