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Cytidine deaminase (CDA) and deoxycytidine monophosphate deaminase (DCTD) play crucial roles in pyrimidine metabolism, affecting DNA synthesis, cell cycle progression, and the efficacy of numerous nucleoside analog-based chemotherapeutics. Given their significance, accurate and sensitive measurement of their enzymatic activity is paramount for both fundamental biochemical research and clinical applications. This review provides a comprehensive overview of the methodologies used to assess CDA and DCTD activity, both established and emerging. We systematically categorize and discuss various approaches, including spectrophotometric, fluorimetric, liquid chromatography-based (Ultraviolet-Visible, fluorescence, and mass spectrometry), radiometric, and cell-based assays. For each method, we present its underlying principles, advantages, and limitations. Furthermore, we draw comparisons across the techniques to highlight their suitability for specific research questions.
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http://dx.doi.org/10.3390/ijms26168045 | DOI Listing |
Int J Mol Sci
August 2025
Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry and Czech Advanced Technology and Research Institute, Palacký University Olomouc, Hněvotínská 5, 779 00 Olomouc, Czech Republic.
Cytidine deaminase (CDA) and deoxycytidine monophosphate deaminase (DCTD) play crucial roles in pyrimidine metabolism, affecting DNA synthesis, cell cycle progression, and the efficacy of numerous nucleoside analog-based chemotherapeutics. Given their significance, accurate and sensitive measurement of their enzymatic activity is paramount for both fundamental biochemical research and clinical applications. This review provides a comprehensive overview of the methodologies used to assess CDA and DCTD activity, both established and emerging.
View Article and Find Full Text PDFProc Natl Acad Sci U S A
June 2025
Institut de Recherches Cliniques de Montréal, Montréal, QC H2W 1R7, Canada.
Cancer therapy is limited by resistance to standard-of-care chemotherapeutic and/or by treatment-associated toxicity. Identifying molecular mechanisms that modulate cellular toxicity is crucial for enhancing treatment efficacy. We characterize CDADC1, a vertebrate-specific orphan enzyme, as an unprecedented eukaryotic dCTP deaminase.
View Article and Find Full Text PDFDalton Trans
June 2025
Key Laboratory of Cluster Science of Ministry of Education, School of Chemistry and Chemical Engineering, Beijing Institute of Technology, Beijing 100081, P. R. China.
Understanding the coordination geometry of nucleotide mono-, di-, and triphosphates is pivotal for unraveling the intricate relationships between molecular structure and biological function, particularly in metal-ligand interactions and their role in biomolecular recognition. This study investigates the structure of nucleotide-metal polymers and their selective interactions with amino acids, specifically tryptophan (Trp) and tyrosine (Tyr). We synthesized and comprehensively analyzed five coordination polymers of cytidine nucleotides: cytidine monophosphate (CMP), deoxycytidine monophosphate (dCMP), cytidine diphosphate (CDP), and cytidine triphosphate (CTP), which are {[Cu(CMP)(bpa)(HO)](CMP)·3HO} (1), {[Cu(dCMP)(4,4'-bipy)(HO)]·4HO} (2), {[Cu(CDP)(azpy)(HO)]·3HO} (3), {[Cd(CDP)(bpa)(HO)]·8HO} (4), and {[Cu(CTP)(2,2'-bipy)]·2HO} (5), where (3), (4) and (5) mark the first report of CDP and CTP coordination complexes.
View Article and Find Full Text PDFProc Natl Acad Sci U S A
May 2025
Laboratory of Structural Biology, International Institute of Molecular and Cell Biology in Warsaw, Warsaw 02-109, Poland.
Vertebrates have evolved an understudied protein termed CDADC1 (NYD-SP15) that contains an inactive N-terminal and active C-terminal DCTD-like domain. Here, we show that human CDADC1 is a (d)CTP-specific deaminase, with a roughly 2-fold in vitro preference for dCTP over CTP. We determined high-resolution cryo-EM structures of CDADC1 in the absence of substrate and in complex with dCTP and 5-methyl-dCTP.
View Article and Find Full Text PDFNaunyn Schmiedebergs Arch Pharmacol
April 2025
Department of Nutrition and Food Hygiene, School of Public Health, Key Laboratory of Precision Nutrition and Health, Ministry of Education, Harbin Medical University, 194 Xuefu Road, Harbin, 150081, Heilongjiang, China.
Depression is a serious mental disease, and its accompanying abnormal changes in peripheral organs, including the kidney, are easy to be ignored. The metabolic abnormalities of the kidney and other organs will inevitably affect the progress of depression through the circulatory system. Quercetin has attracted much attention as a flavonoid with anti-inflammatory, antioxidant, neuroprotective, and antidepressant potential.
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