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

Stachelska-Wierzchowska, Alicja; Wierzchowski, Jacek; Wielgus‐Kutrowska, Beata; Mikleušević, Goran

doi:10.3390/molecules181012587

Cited by 19 publications

(22 citation statements)

References 36 publications

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“…This ambiguity in the ribosylation sites in enzymatic reactions, catalyzed by PNP, was also previously observed in 8-azapurines [39,40], and probably results from the plasticity of the active site of this class of enzymes. For example, in the X-ray crystal structure of hexameric PNP from H. pylori, bound to an inhibitor formycin A (8-aza-N 9 -deazaadenosine), both the standard (anti) and non-standard (syn) conformations of the inhibitor were revealed [41].…”

Section: Enzymatic Syntheses Of the Tri-cyclic Ribosides And Their Prsupporting

confidence: 65%

Tricyclic Nucleobase Analogs and Their Ribosides as Substrates and Inhibitors of Purine-Nucleoside Phosphorylases III. Aminopurine Derivatives

Stachelska-Wierzchowska

Wierzchowski

Górka

et al. 2020

Molecules

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Etheno-derivatives of 2-aminopurine, 2-aminopurine riboside, and 7-deazaadenosine (tubercidine) were prepared and purified using standard methods. 2-Aminopurine reacted with aqueous chloroacetaldehyde to give two products, both exhibiting substrate activity towards bacterial (E. coli) purine-nucleoside phosphorylase (PNP) in the reverse (synthetic) pathway. The major product of the chemical synthesis, identified as 1,N2-etheno-2-aminopurine, reacted slowly, while the second, minor, but highly fluorescent product, reacted rapidly. NMR analysis allowed identification of the minor product as N2,3-etheno-2-aminopurine, and its ribosylation product as N2,3-etheno-2-aminopurine-N2-β-d-riboside. Ribosylation of 1,N2-etheno-2-aminopurine led to analogous N2-β-d-riboside of this base. Both enzymatically produced ribosides were readily phosphorolysed by bacterial PNP to the respective bases. The reaction of 2-aminopurine-N9-β -d-riboside with chloroacetaldehyde gave one major product, clearly distinct from that obtained from the enzymatic synthesis, which was not a substrate for PNP. A tri-cyclic 7-deazaadenosine (tubercidine) derivative was prepared in an analogous way and shown to be an effective inhibitor of the E. coli, but not of the mammalian enzyme. Fluorescent complexes of amino-purine analogs with E. coli PNP were observed.

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Section: Enzymatic Syntheses Of the Tri-cyclic Ribosides And Their Prsupporting

confidence: 65%

Tricyclic Nucleobase Analogs and Their Ribosides as Substrates and Inhibitors of Purine-Nucleoside Phosphorylases III. Aminopurine Derivatives

Stachelska-Wierzchowska

Wierzchowski

Górka

et al. 2020

Molecules

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“…[72] Ribosides and deoxyribosides of 8-azaDaPur have been synthesized many years ago, [73,74] but their fluorescence properties were not examined until recently. We have found that enzymatically generated N9-, N8-, and N7-β-d-ribosides of 8-azaDaPur are highly fluorescent, [75] with yields of ca. 0.9, 0.4, and 0.06, respectively.…”

Section: 6-diamino-8-azapurine and The Corresponding Ribosidesmentioning

confidence: 99%

“…It is evident that some of the described compounds may be classified as "super photoacids." [34] It was found that many of the presented 8-azapurine analogs and their ribosides, including the nontypical N7-and N-8-β-d-ribosides, are accepted as substrates by enzymes of nucleoside metabolism, [75,76] and therefore can be applied as fluorescent probes of these systems.…”

Section: 6-diamino-8-azapurine and The Corresponding Ribosidesmentioning

confidence: 99%

Excited-State Proton Transfer and Phototautomerism in Nucleobase and Nucleoside Analogs: A Mini-Review

Wierzchowski¹

2014

Nucleosides, Nucleotides and Nucleic Acids

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Intermolecular excited-state proton transfer (ESPT) has been observed in several fluorescent nucleobase and/or nucleoside analogs. In the present work, some new examples of ESPT in this class of compounds are presented together with a brief recapitulation of the previously published data. The nucleobases, nucleosides, and their analogs contain many basic and acidic centers and therefore their ESPT behavior may be complex. To interpret the complex data, it is usually necessary to determine the microscopic pK* values for each (or most) of the possible ESPT centers. Typical approach to solve this problem is by analysis of the alkyl derivatives, in which the possibility of the ESPT is reduced. Of particular interest are examples of "phototautomerization via the cation," observed in several systems, which in the neutral media do not undergo ESPT. Protonation of the molecule in the ground state facilitates the two-step phototautomerism in several systems, including formycin A and 2-amino-8-azadenine. Fluorescence of the nucleobase and nucleoside analogs undergoing ESPT is usually solvent-, isotope-, and buffer-ion sensitive, and in some systems the ESPT can be promoted by environmental factors, e.g., the presence of buffer ions. This sensitivity to the microenvironment parameters makes the ESPT systems potentially useful for biological applications.

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“…It has been shown previously, that enzymatic ribosylation of some nucleobase analogs with PNP as a biocatalyst leads to non-typical ribosides, with ribose moiety attached not necessarily to purine N9, but also to other nitrogen atoms (Stachelska-Wierzchowska et al, 2013;Stachelska-Wierzchowska et al, 2016;Stachelska-Wierzchowska et al, 2018;Stachelska-Wierzchowska et al, 2019). In particular, while the E. coli PNP directs the ribosyl group predominantly to the N9 of ethenoadenine, the calf enzyme leads to almost exclusively N 6riboside (Stachelska-Wierzchowska et al, 2018).…”

Section: Resultsmentioning

confidence: 99%

Fluorescence of the tri-cyclic adenine and isoguanine derivatives and their ribosides: possible analytical applications

Stachelska-Wierzchowska

Wierzchowski

2019

Acta Biochim Pol

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Fluorescent tri-cyclic purine analogs, derivatives of isoguanine and adenine, were examined as potential substrates of purine-nucleoside phosphorylase. It was found previously that etheno- derivatives of both compounds are ribosylated in phosphate-free media, but ribosylation places in some instances differ from purine N9. New ribosides are examined as potential substrates of human blood PNP and indicators of this enzyme. Of these, N6-riboside of 1,N6-etheno-adenine was found the most promising.

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

Cited by 19 publications

References 36 publications

Tricyclic Nucleobase Analogs and Their Ribosides as Substrates and Inhibitors of Purine-Nucleoside Phosphorylases III. Aminopurine Derivatives

Tricyclic Nucleobase Analogs and Their Ribosides as Substrates and Inhibitors of Purine-Nucleoside Phosphorylases III. Aminopurine Derivatives

Excited-State Proton Transfer and Phototautomerism in Nucleobase and Nucleoside Analogs: A Mini-Review

Fluorescence of the tri-cyclic adenine and isoguanine derivatives and their ribosides: possible analytical applications

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