Nucleoside and deoxynucleoside phosphorylation in formamide solutions

Abstract

Nucleosides or deoxynucleosides were converted to a number of phosphorylated nucleotide and deoxynucleotide derivatives by ammonium of alkali dihydrogen phosphates in formamide. Conversions were smaller and slower at room temperature and greater and faster at elevated temperatures. Nucleotides afforded product mixtures to those obtained for nucleosides under the same conditions, indicating the occurrence of transphosphorylation processes. Products of the reaction at elevated temperatures were cyclic nucleotides, nucleoside monophosphates, nucleoside diphosphates and cyclic nucleotide phosphates. The relative amounts of products formed were quite temperature dependent. Cyclic nucleotides were found to be in greatest abundance for reactions run at 125 degree or above. Relative yields of 2',3' and 5' nucleotides and 3' and 5' deoxynucleotides from several experiments are reported. 5'Monophosphates were generally found to be present in larger quantities than 2' or 3' monophosphates. 2'-Deoxyadenosine showed a preference for phosphorylation at the 3' position. Conclusion reached from mechanistic studies are that the phosphorylations are a series of equilibrium reactions, with cyclic nucleotides being formed irreversibly.