Two fluorogenic substrates for purine nucleoside phosphorylase, selective for mammalian and bacterial forms of the enzyme

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

doi:10.1016/j.ab.2013.10.017

Cited by 11 publications

(22 citation statements)

References 13 publications

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“…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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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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“…Implemented into the workflow of a high-throughput assay, this methodology facilitated a > 20-fold reduction of data acquisition time and a roughly fivefold decrease in cost compared to conventional HPLC, while maintaining very comparable accuracy and excellent reproducibility. Unlike other non-HPLC-based methods, [15][16][17][18][19][20][21][22][23][24][25][26][27] our approach offers a uniquely broad substrate spectrum, including all natural and several examples of modified nucleosides, as well as high adaptability and the straightforward application to any substrate without the need for laborious method development.…”

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confidence: 99%

Spectral Unmixing‐Based Reaction Monitoring of Transformations between Nucleosides and Nucleobases

et al. 2020

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The increased interest in (enzymatic) transformations between nucleosides and nucleobases has demanded the development of efficient analytical tools. In this report, we present an update and extension of our recently described method for monitoring these reactions by spectral unmixing. The presented method uses differences in the UV absorption spectra of nucleosides and nucleobases after alkaline quenching to derive their ratio based on spectral shape by fitting normalized reference spectra. It is applicable to a broad compound spectrum comprising more than 35 examples, offers HPLC‐like accuracy, ease of handling and significant reductions in both cost and data acquisition time compared to other methods. This contribution details the principle of monitoring reactions by spectral unmixing, gives recommendations regarding solutions to common problems and applications that necessitate special sample treatment. We provide software, workflows and reference spectra that facilitate the straightforward and versatile application of the method.

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“…The purpose of the present paper is examination of substrate properties of new fluorescent ribosides toward the human blood enzyme, and their analytical potential in clinical investigations. We have previously identified two such substrates, isomeric ribosides of 2,6-diamino-8-azapurine (Wierzchowski et al, 2014), and now extend our work to include tri-cyclic ribosides with different spectral characteristics.…”

Section: Introductionmentioning

confidence: 71%

“…We have shown previously that some phosphorolytic reactions are easily observed spectrally or fluorimetrically using 1000-fold diluted whole blood lysates as a catalyst in ca. 50 mM phosphate buffer (Wierzchowski et al, 2002;Wierzchowski et al, 2014). We have therefore examined the new ribosides as potential substrates for the human PNP, using the same methodology (except optical pathlength of the cuvettes, reduced now to 4 mm).…”

Section: Resultsmentioning

confidence: 99%

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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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Two fluorogenic substrates for purine nucleoside phosphorylase, selective for mammalian and bacterial forms of the enzyme

Cited by 11 publications

References 13 publications

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

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

Spectral Unmixing‐Based Reaction Monitoring of Transformations between Nucleosides and Nucleobases

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

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