Cargando…

Site-Selective Ribosylation of Fluorescent Nucleobase Analogs Using Purine-Nucleoside Phosphorylase as a Catalyst: Effects of Point Mutations

Enzymatic ribosylation of fluorescent 8-azapurine derivatives, like 8-azaguanine and 2,6-diamino-8-azapurine, with purine-nucleoside phosphorylase (PNP) as a catalyst, leads to N9, N8, and N7-ribosides. The final proportion of the products may be modulated by point mutations in the enzyme active sit...

Descripción completa

Detalles Bibliográficos
Autores principales: Stachelska-Wierzchowska, Alicja, Wierzchowski, Jacek, Bzowska, Agnieszka, Wielgus-Kutrowska, Beata
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6274182/
https://www.ncbi.nlm.nih.gov/pubmed/26729076
http://dx.doi.org/10.3390/molecules21010044
_version_ 1783377559719772160
author Stachelska-Wierzchowska, Alicja
Wierzchowski, Jacek
Bzowska, Agnieszka
Wielgus-Kutrowska, Beata
author_facet Stachelska-Wierzchowska, Alicja
Wierzchowski, Jacek
Bzowska, Agnieszka
Wielgus-Kutrowska, Beata
author_sort Stachelska-Wierzchowska, Alicja
collection PubMed
description Enzymatic ribosylation of fluorescent 8-azapurine derivatives, like 8-azaguanine and 2,6-diamino-8-azapurine, with purine-nucleoside phosphorylase (PNP) as a catalyst, leads to N9, N8, and N7-ribosides. The final proportion of the products may be modulated by point mutations in the enzyme active site. As an example, ribosylation of the latter substrate by wild-type calf PNP gives N7- and N8-ribosides, while the N243D mutant directs the ribosyl substitution at N9- and N7-positions. The same mutant allows synthesis of the fluorescent N7-β-d-ribosyl-8-azaguanine. The mutated form of the E. coli PNP, D204N, can be utilized to obtain non-typical ribosides of 8-azaadenine and 2,6-diamino-8-azapurine as well. The N7- and N8-ribosides of the 8-azapurines can be analytically useful, as illustrated by N7-β-d-ribosyl-2,6-diamino-8-azapurine, which is a good fluorogenic substrate for mammalian forms of PNP, including human blood PNP, while the N8-riboside is selective to the E. coli enzyme.
format Online
Article
Text
id pubmed-6274182
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-62741822018-12-28 Site-Selective Ribosylation of Fluorescent Nucleobase Analogs Using Purine-Nucleoside Phosphorylase as a Catalyst: Effects of Point Mutations Stachelska-Wierzchowska, Alicja Wierzchowski, Jacek Bzowska, Agnieszka Wielgus-Kutrowska, Beata Molecules Article Enzymatic ribosylation of fluorescent 8-azapurine derivatives, like 8-azaguanine and 2,6-diamino-8-azapurine, with purine-nucleoside phosphorylase (PNP) as a catalyst, leads to N9, N8, and N7-ribosides. The final proportion of the products may be modulated by point mutations in the enzyme active site. As an example, ribosylation of the latter substrate by wild-type calf PNP gives N7- and N8-ribosides, while the N243D mutant directs the ribosyl substitution at N9- and N7-positions. The same mutant allows synthesis of the fluorescent N7-β-d-ribosyl-8-azaguanine. The mutated form of the E. coli PNP, D204N, can be utilized to obtain non-typical ribosides of 8-azaadenine and 2,6-diamino-8-azapurine as well. The N7- and N8-ribosides of the 8-azapurines can be analytically useful, as illustrated by N7-β-d-ribosyl-2,6-diamino-8-azapurine, which is a good fluorogenic substrate for mammalian forms of PNP, including human blood PNP, while the N8-riboside is selective to the E. coli enzyme. MDPI 2015-12-28 /pmc/articles/PMC6274182/ /pubmed/26729076 http://dx.doi.org/10.3390/molecules21010044 Text en © 2015 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons by Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Stachelska-Wierzchowska, Alicja
Wierzchowski, Jacek
Bzowska, Agnieszka
Wielgus-Kutrowska, Beata
Site-Selective Ribosylation of Fluorescent Nucleobase Analogs Using Purine-Nucleoside Phosphorylase as a Catalyst: Effects of Point Mutations
title Site-Selective Ribosylation of Fluorescent Nucleobase Analogs Using Purine-Nucleoside Phosphorylase as a Catalyst: Effects of Point Mutations
title_full Site-Selective Ribosylation of Fluorescent Nucleobase Analogs Using Purine-Nucleoside Phosphorylase as a Catalyst: Effects of Point Mutations
title_fullStr Site-Selective Ribosylation of Fluorescent Nucleobase Analogs Using Purine-Nucleoside Phosphorylase as a Catalyst: Effects of Point Mutations
title_full_unstemmed Site-Selective Ribosylation of Fluorescent Nucleobase Analogs Using Purine-Nucleoside Phosphorylase as a Catalyst: Effects of Point Mutations
title_short Site-Selective Ribosylation of Fluorescent Nucleobase Analogs Using Purine-Nucleoside Phosphorylase as a Catalyst: Effects of Point Mutations
title_sort site-selective ribosylation of fluorescent nucleobase analogs using purine-nucleoside phosphorylase as a catalyst: effects of point mutations
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6274182/
https://www.ncbi.nlm.nih.gov/pubmed/26729076
http://dx.doi.org/10.3390/molecules21010044
work_keys_str_mv AT stachelskawierzchowskaalicja siteselectiveribosylationoffluorescentnucleobaseanalogsusingpurinenucleosidephosphorylaseasacatalysteffectsofpointmutations
AT wierzchowskijacek siteselectiveribosylationoffluorescentnucleobaseanalogsusingpurinenucleosidephosphorylaseasacatalysteffectsofpointmutations
AT bzowskaagnieszka siteselectiveribosylationoffluorescentnucleobaseanalogsusingpurinenucleosidephosphorylaseasacatalysteffectsofpointmutations
AT wielguskutrowskabeata siteselectiveribosylationoffluorescentnucleobaseanalogsusingpurinenucleosidephosphorylaseasacatalysteffectsofpointmutations