Design and synthesis of double‐stranded oligonucleotides containing reactive acylphosphate internucleotide groups

Abstract

DNA duplex and dumbbells containing chemically active acylphosphate internucleotide groups were synthesized. To obtain these compounds the chemical ligation method was used. The acylphosphate group was inserted into a DNA duplex and dumbbells as a result of template-directed condensation of 5P-phosphate and especially introduced 3P-carboxy groups of oligonucleotides. 1-Ethyl-3-(3P-dimethylaminopropyl)carbodiimide (EDC) was used as a condensing agent. Oligonucleotides containing a carboxy group were obtained by the interaction of their 3P-phosphate with glycine methyl ester under the action of EDC, followed by ester hydrolysis. The yields of acylphosphatecontaining double-stranded oligonucleotides achieved 15^25% depending on the structure of their precursors. It was shown that these compounds are acylating agents and are efficiently cleaved in near-physiological conditions under the action of ethylenediamine or N-methylimidazole. These results indicate that doublestranded oligonucleotides carrying acylphosphate internucleotide groups could constitute new crosslinking reagents for affinity modification of DNA recognizing proteins.z 1998 Federation of European Biochemical Societies.

Functional Interrogation of the Kinome Using Nucleotide Acyl Phosphates

“…Methyl acetyl phosphate is known to inactivate several enzymes by acylation (22). The electrophilic properties of acyl phosphates have also been used in a number of applications such as cross-linking reagents for hemoglobins (23) and the affinity modification of DNA-recognizing proteins (24). Little research has been reported for acyl phosphates of di-and triphosphates.…”