1,N6-ethenoadenine and other Fluorescent Nucleobase Analogs as Substrates for Purine-Nucleoside Phosphorylases: Spectroscopic and Kinetic Studies

Author(s): Jacek Wierzchowski* , Alicja Stachelska-Wierzchowska , Beata Wielgus-Kutrowska , Agnieszka Bzowska .

Journal Name: Current Pharmaceutical Design

Volume 23 , Issue 45 , 2017

Abstract:

Background: Purine-nucleoside phosphorylase (PNP) is known as a tool for the synthesis of various nucleosides and nucleoside analogues. Mechanism, properties, molecular diversity and inhibitors of PNP, particularly these of pharmacological significance, are briefly characterized.

Methods: UV and fluorescence spectroscopy was used for kinetic experiments, and HPLC chromatography for product analyses.

Results: Applications of various forms of PNP to synthesis of selected fluorescent nucleosides, particularly ribosides of 1,N6-ethenoadenine and various 8-azapurines (triazolo[4,5-d]pyrimidines) are reviewed. Different specificity of various PNP forms is described towards nucleobase and analog substrates as well as variable ribosylation sites observed in some reactions, with a possibility to further modify these features via the site-directed mutagenesis.

Conclusion: Present and future applications of the fluorescent or fluorogenic ribosides are discussed, with particular emphasis on biochemical and clinical analyses with improved sensitivity.

Keywords: Purine-nucleoside phosphorylase, isozymes, nucleoside analogs, fluorescence, enzyme assays, enzyme-ligand complexes.

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Article Details

VOLUME: 23
ISSUE: 45
Year: 2017
Page: [6948 - 6966]
Pages: 19
DOI: 10.2174/1381612823666171011103551
Price: $58

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