Cytidine and dCMP Deaminases—Current Methods of Activity Analysis

• Published in: International journal of molecular sciences Vol. 26; no. 16; p. 8045
• Main Authors: Ligasová, Anna, Horejšová, Martina, Brumarová, Radana, Friedecký, David, Koberna, Karel
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 20.08.2025; MDPI
• Subjects: Ammonia; Analysis; Animals; Catalysis; Cell cycle; Chromatography, Liquid; Cytidine - metabolism; Cytidine Deaminase - metabolism; DNA synthesis; Enzyme Assays - methods; Enzymes; Fluorescence; Genes; Humans; Liquid chromatography; Mass Spectrometry; Metabolism; Metabolites; Nucleosides; Pharmacokinetics; Physiological aspects; Polymorphism; Pyrimidines; Review; Czech Republic; spectrophotometry; cell-based approaches; modified nucleosides; LC-MS; cytidine deaminase; dCMP deaminase
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms26168045

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.