Enzymatic Synthesis of Bis(5′-Nucleosidyl) Tetra- and Triphosphates

The total fraction of aminoacyl-tRNA synthases from Escherichia coli has been shown to catalyze the synthesis of the bis(5'-nucleosidyl) oligophosphates Ap sub(4)AZT, Ap sub(4)d4T, Ap sub(4)3TC, and Ap sub(4)ACV, as well as Ap sub(3)AZT and Ap sub(3)d4T, from [ alpha - super(32)P]ATP and the corresponding nucleoside-5'-tri(or di)phosphate. The resulting compounds, characterized by HPLC, are resistant to alkaline phosphatase. Ap sub(4)AZT, Ap sub(4)d4T, and Ap sub(4)3TC are formed with approximately equal efficiency, whereas the efficiencies of the synthesis of Ap sub(4)ACV, Ap sub(3)AZT, and Ap sub(3)d4T are three- to fivefold lower.