Next Article in Journal
Comparative Analysis of Click Chemistry Mediated Activity-Based Protein Profiling in Cell Lysates
Next Article in Special Issue
An Efficient and Facile Methodology for Bromination of Pyrimidine and Purine Nucleosides with Sodium Monobromoisocyanurate (SMBI)
Previous Article in Journal
Flavonoid Profile of Saskatoon Berries (Amelanchier alnifolia Nutt.) and Their Health Promoting Effects
Previous Article in Special Issue
Glycosyl-Nucleolipids as New Bioinspired Amphiphiles
Molecules 2013, 18(10), 12587-12598; doi:10.3390/molecules181012587
Article

Enzymatic Synthesis of Highly Fluorescent 8-Azapurine Ribosides Using a Purine Nucleoside Phosphorylase Reverse Reaction: Variable Ribosylation Sites

1,* , 1
, 2
 and 3
1 Department of Biophysics, University of Varmia & Masuria, 4 Oczapowskiego St., 10-719 Olsztyn, Poland 2 Division of Biophysics, Institute of Experimental Physics, University of Warsaw, Zwirki i Wigury 93, 02-089 Warsaw, Poland 3 Division of Physical Chemistry, Rudjer Bošković Institute, POB 180, HR-10002 Zagreb, Croatia
* Author to whom correspondence should be addressed.
Received: 30 August 2013 / Revised: 25 September 2013 / Accepted: 30 September 2013 / Published: 11 October 2013
(This article belongs to the Special Issue Synthesis of Nucleosides, Nucleotides and Their Derivatives)
Download PDF [414 KB, 18 June 2014; original version 18 June 2014]

Abstract

Various forms of purine-nucleoside phosphorylase (PNP) were used as catalysts of enzymatic ribosylation of selected fluorescent 8-azapurines. It was found that the recombinant calf PNP catalyzes ribosylation of 2,6-diamino-8-azapurine in a phosphate-free medium, with ribose-1-phosphate as ribose donor, but the ribosylation site is predominantly N7 and N8, with the proportion of N8/N7 ribosylated products markedly dependent on the reaction conditions. Both products are fluorescent. Application of the E. coli PNP gave a mixture of N8 and N9-substituted ribosides. Fluorescence of the ribosylated 2,6-diamino-8-azapurine has been briefly characterized. The highest quantum yield, ~0.9, was obtained for N9-β-d-riboside (λmax 365 nm), while for N8-β-d-riboside, emitting at ~430 nm, the fluorescence quantum yield was found to be close to 0.4. Ribosylation of 8-azaguanine with calf PNP as a catalyst goes exclusively to N9. By contrast, the E. coli PNP ribosylates 8-azaGua predominantly at N9, with minor, but highly fluorescent products ribosylated at N8/N7.
Keywords: 8-azapurines; nucleosides; enzymatic synthesis; fluorescence; purine-nucleoside phosphorylase 8-azapurines; nucleosides; enzymatic synthesis; fluorescence; purine-nucleoside phosphorylase
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Share & Cite This Article

Export to BibTeX |
EndNote


MDPI and ACS Style

Stachelska-Wierzchowska, A.; Wierzchowski, J.; Wielgus-Kutrowska, B.; Mikleušević, G. Enzymatic Synthesis of Highly Fluorescent 8-Azapurine Ribosides Using a Purine Nucleoside Phosphorylase Reverse Reaction: Variable Ribosylation Sites. Molecules 2013, 18, 12587-12598.

View more citation formats

Related Articles

Article Metrics

Comments

Citing Articles

[Return to top]
Molecules EISSN 1420-3049 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert