Enzymatic synthesis of 5-azacytidine 5′-triphosphate from 5-azacytidine

✍ Scribed by Thomas T. Lee; Richard L. Momparler

• Publisher: Elsevier Science
• Year: 1976
• Tongue: English
• Weight: 468 KB
• Volume: 71
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(76)90011-7

✦ Synopsis

Azacytidine 5'-monophosphate (S-aza-CMP) was synthesized enzymatically from 5-azacytidine (S-aza-C) in a reaction catalyzed by uridine-cytidine kinase. In a second step, 5-azacytidine 5'-triphosphate (5-aza-CTP) was synthesized enzymatically from 5-aza-CMP using CMP kinase and nucleoside diphosphokinase. Due to the chemical instability of the triazide ring of 5-azacytosine at neutral and alkaline pH, the enzymatic synthesis and purification of the nucleotides by ion exchange chromatography were performed at acid pH. The enzymatically synthesized 5-aza-CTP had an ultraviolet absorbance spectrum at pH 5.5 similar to the spectrum of 5-aza-C. In the DNA-dependent RNA polymerase reaction, 5-aza-CTP inhibited the incorporation of [3H]CTP, but not [3H]UTP, into RNA.