Abstract
3′-Deoxy-3′,4′-didehydro-cytidine triphosphate (ddhCTP) is a novel antiviral molecule produced by the enzyme viperin during the early stages of the innate immune response. ddhCTP has been shown to act as a chain terminator of flavivirus RNA-dependent RNA polymerases. To date, synthesis of ddhCTP requires complicated synthetic protocols or isolation of the enzyme viperin to catalyze the production of ddhCTP from CTP. Recombinant viperin approaches preclude the production of highly pure ddhCTP (free of contaminants such as CTP), whereas the chemical synthesis involves techniques or equipment not readily available to most laboratories. Herein, we describe the chemoenzymatic synthesis of ddhCTP, starting from commercially available ddhC. We utilize these methods to produce milligram quantities of ddhCTP, ddhCDP, and ddhCMP. Using purified semisynthetic ddhCTP and fully synthetic ddhCTP, we also show ddhCTP does not inhibit NAD+-dependent enzymes such as glyceraldehyde 3-phosphate dehydrogenase, malate dehydrogenase, or lactate dehydrogenase, contrary to a recent report.
Original language | English (US) |
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Pages (from-to) | 322-326 |
Number of pages | 5 |
Journal | ACS Bio and Med Chem Au |
Volume | 3 |
Issue number | 4 |
DOIs | |
State | Published - Aug 16 2023 |
Keywords
- GAPDH
- Viperin
- anion exchange chromatography
- chemoenzymatic synthesis
- ddhCTP
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology
- Pharmaceutical Science
- Drug Discovery