Determination of pyrimidine deoxynucleoside triphosphates in leukaemia cell extracts containing 1-beta-D-arabinofuranosylcytosine triphosphate.
Deoxynucleoside 5'-triphosphates (dNTPs) can be determined in cell extracts by high performance liquid chromatography after prior selective degradation of ribonucleoside 5'-triphosphates with sodium periodate and methylamine. When the method is used for the evaluation of deoxynucleoside triphosphates in 1-beta-D-arabinofuranosylcytosine triphosphate (ara-CTP)-containing cell extracts, an additional peak coeluting with thymidine triphosphate (dTTP) is observed. This peak is due to the formation of a carboxylic acid derivative of ara-CTP by periodate oxidation, and it can lead to considerable overestimation of dTTP. Formation of this peak can be avoided by using alkaline reaction conditions (pH 7.5) and by changing the sequence of addition of the reagents used in the periodation procedure. By employing this modified protocol, cellular dNTP and ara-CTP levels can be monitored in extracts of leukaemic blasts during cytosine arabinoside treatment in two separate HPLC runs.
Chiba
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P
Department of Medical Chemistry, University of Vienna, Austria.
Szekeres
T
T
Jäger
W
W
D000255:Adenosine Triphosphate / Q000032:analysis; D001085:Arabinofuranosylcytosine Triphosphate / Q000032:analysis*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D003570:Cytidine Triphosphate / Q000032:analysis; D003854:Deoxyribonucleotides / Q000032:analysis*; D006160:Guanosine Triphosphate / Q000032:analysis; D006801:Humans; D007938:Leukemia; D012265:Ribonucleotides / Q000032:analysis*; D013942:Thymine Nucleotides / Q000032:analysis*; D014544:Uridine Triphosphate / Q000032:analysis
D003854:Deoxyribonucleotides; D012265:Ribonucleotides; D013942:Thymine Nucleotides; D001085:Arabinofuranosylcytosine Triphosphate; D003570:Cytidine Triphosphate; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; C024157:thymidine 5'-triphosphate; D014544:Uridine Triphosphate