F-ara-AMP is a substrate of cytoplasmic 5'-nucleotidase II (cN-II): HPLC and NMR studies of enzymatic dephosphorylation.
Intracellular accumulation of triphosphorylated derivatives is essential for the cytotoxic activity of nucleoside analogues. Different mechanisms opposing this accumulation have been described. We have investigated the dephosphorylation of monophosphorylated fludarabine (F-ara-AMP) by the purified cytoplasmic 5'-nucleotidase cN-II using HPLC and NMR. These studies clearly showed that cN-II was able to convert F-ara-AMP into its non phosphorylated form, F-ara-A, with a Km in the millimolar range and Vmax = 35 nmol/min/mg, with both methods. Cytoplasmic 5'-nucleotidase cN-II can degrade this clinically useful cytotoxic nucleoside analogue and its overexpression is thus likely to be involved in resistance to this compound.
Jordheim
Lars P
LP
INSERM U590, Laboratoire de Cytologie Analytique, Faculté de Médecine Rockefeller, Université Claude Bernard Lyon I, Lyon, France. jordheim@yahoo.com
Cros
Emeline
E
Galmarini
Carlos M
CM
Dumontet
Charles
C
Bretonnet
Anne-Sophie
AS
Krimm
Isabelle
I
Lancelin
Jean-Marc
JM
Gagnieu
Marie-Claude
MC
D015720:5'-Nucleotidase / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*; D002851:Chromatography, High Pressure Liquid; D003593:Cytoplasm / Q000201:enzymology*; D006801:Humans; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D014740:Vidarabine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D001084:Vidarabine Phosphate / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism
D001084:Vidarabine Phosphate; C042382:fludarabine phosphate; D015720:5'-Nucleotidase; C508048:NT5C2 protein, human; D014740:Vidarabine; C024352:fludarabine