ArticleMethods and protocols2019
A UV/Vis Spectroscopy-Based Assay for Monitoring of Transformations Between Nucleosides and Nucleobases.
Article in Methods and protocols, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed.
- Water Soluble-Nitrogenous Secondary Metabolites From Talaromyces annesophieae MD2 Exhibit Anti-Bacterial, Anti-Cancer, Anti-Oxidant, and Cytoprotective Activities.MicrobiologyOpen · 2026Article
- Interactions of Nedaplatin with Nucleobases and Purine Alkaloids: Their Role in Cancer Therapy.Biomedicines · 2025Article
- Synthesis and Characterization of NUC-7738, an Aryloxy Phosphoramidate of 3'-Deoxyadenosine, as a Potential Anticancer Agent.Journal of medicinal chemistry · 2022Article
- Thermostable adenosine 5'-monophosphate phosphorylase from Thermococcus kodakarensis forms catalytically active inclusion bodies.Scientific reports · 2021Article
- Optimized Biocatalytic Synthesis of 2-Selenopyrimidine Nucleosides by Transglycosylation*.Chembiochem : a European journal of chemical biology · 2021Article
- The Peculiar Case of the Hyper-thermostable Pyrimidine Nucleoside Phosphorylase from Thermus thermophilus*.Chembiochem : a European journal of chemical biology · 2021Article
- Kinetic Analysis of the Hydrolysis of Pentose-1-phosphates through Apparent Nucleoside Phosphorolysis Equilibrium Shifts*.Chemphyschem : a European journal of chemical physics and physical chemistry · 2021Article
- Spectral Unmixing-Based Reaction Monitoring of Transformations between Nucleosides and Nucleobases.Chembiochem : a European journal of chemical biology · 2020Article
- General Principles for Yield Optimization of Nucleoside Phosphorylase-Catalyzed Transglycosylations.Chembiochem : a European journal of chemical biology · 2020Article
- Efficient Biocatalytic Synthesis of Dihalogenated Purine Nucleoside Analogues Applying Thermodynamic Calculations.Molecules (Basel, Switzerland) · 2020Article
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6 authors.
Funding
Abstract
Efficient reaction monitoring is crucial for data acquisition in kinetic and mechanistic studies. However, for conversions of nucleosides to their corresponding nucleobases, as observed in enzymatically catalyzed nucleoside phosphorylation reactions, the current analytical arsenal does not meet modern requirements regarding cost, speed of analysis and high throughput. Herein, we present a UV/Vis spectroscopy-based assay employing an algorithm for spectral unmixing in a 96-well plate format. The algorithm relies on fitting of reference spectra of nucleosides and their bases to experimental spectra and allows determination of nucleoside/nucleobase ratios in solution with high precision. The experimental procedure includes appropriate dilution of a sample into aqueous alkaline solution, transfer to a multi-well plate, measurement of a UV/Vis spectrum and subsequent in silico spectral unmixing. This enables data collection in a high-throughput fashion and reduces costs compared to state-of-the-art HPLC analyses by approximately 5-fold while being 20-fold faster and offering comparable precision. Additionally, the method is robust regarding dilution and sample transfer errors as it only considers spectral form and not absolute intensity. It can be applied to all natural nucleosides and nucleobases and even unnatural ones as demonstrated by several examples.
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