Synthesis of 8-(2-4 dinitrophenyl 2-6 aminohexyl) amino-adenosine 5' triphosphate: biological properties and potential uses.
We have synthetised 8-(2-4 dinitrophenyl 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 recognized by specific antibodies which can then be detected by anti-antibodies coupled to an enzyme. DNP 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 labelling as a substitute to the radioactive detection of nucleic acids, are discussed.
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D000936:Antigen-Antibody Complex; D004253:DNA Nucleotidylexotransferase / Q000378:metabolism*; D004254:DNA Nucleotidyltransferases / Q000378:metabolism*; D004256:DNA Polymerase I / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D007106:Immune Sera; D007700:Kinetics; D010957:Plasmids
D000936:Antigen-Antibody Complex; D007106:Immune Sera; C036874:8-(2,4-dinitrophenyl-2,6-aminohexyl)aminoadenosine 5'-triphosphate; D000255:Adenosine Triphosphate; D004254:DNA Nucleotidyltransferases; D004253:DNA Nucleotidylexotransferase; D004256:DNA Polymerase I; D004259:DNA-Directed DNA Polymerase