Synthesis of 8-(2-4 dinitrophenyl 2-6 aminohexyl) amino–adenosine 5′ triphosphate: biological properties and potential uses

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Abstract
We have synthetised 8–(2–4 dinitrohenyl 2–6 aminohexyl) amino–adenosine 5′ triphosphate (in short: rATP–DNP), a derivative of ATP which carries a dinitrophenyl group. We show that rATP–DNP is a substrate for calf thymus deoxynucleotidyl terminal transferase (EC 2.7.7.31) and E. coli DNA polymerase I (Kornberg polymerase EC 2.7.7.7). It can therefore be incorporated into DNA molecules by elongation from 3′ ends or by nick translation. The incorporated dinitrophenyl group can be reconginzed by specific antibodies which can then be detected by anti-antibodies coupled to an enzyme. ENP groups could also be introduced into DNA after enzymatic incorporation of 8–aminohexyl adenosine 5′ triphosphate and reaction with 1–fluoro–2–4–dinitrobenzene. Thus, DNA molecules carrying DNP groups can ultimately be revealed by enzymatic coloured reactions. Potential uses of this enzymatic labeling as a substitute to the radioactive detection of nucleic acids, are discussed.