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

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

Volume: 23 Issue: 45

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

Affiliation:

• Department of Biophysics, Faculty of Food Sciences, University of Varmia & Masuria, Olsztyn,Poland

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

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.

Cite this article as:

Wierzchowski Jacek *, Stachelska-Wierzchowska Alicja, Wielgus-Kutrowska Beata and Bzowska Agnieszka, 1,N6-ethenoadenine and other Fluorescent Nucleobase Analogs as Substrates for Purine-Nucleoside Phosphorylases: Spectroscopic and Kinetic Studies, Current Pharmaceutical Design 2017; 23 (45) . https://dx.doi.org/10.2174/1381612823666171011103551