Thymidine phosphorylase is noncompetitively inhibited by 5'-O-trityl-inosine (KIN59) and related compounds.
We found that 5'-O-trityl-inosine (KIN59) inhibits recombinant bacterial (E. coli) and human thymidine phosphorylase (TPase) with an IC50 of 44 microM and 67 microM, respectively. In contrast to previously described TPase inhibitors, KIN59 does not compete with thymidine (dThd) at the pyrimidine nucleoside-binding site or with inorganic phosphate (Pi) at the phosphate-binding site of the enzyme. These findings are strongly suggestive for the presence of an allosteric binding site at the enzyme. TPase is identical to the angiogenic protein platelet-derived endothelial cell growth factor (PD-ECGF). As such, PD-ECGF stimulates angiogenesis in the chick chorioallantoic membrane (CAM) assay. This angiogenic response was completely inhibited by KIN59. Inosine did not inhibit the enzyme or the angiogenic effect of TPase, confirming that the 5'-O-trityl group in KIN59 is essential for the observed effect. Our observations indicate that allosteric sites in TPase may regulate its biological activity.
Liekens
S
S
Rega Institute for Medical Research, KU Leuven, Leuven, Belgium. sandra.liekens@rega.kuleuven.be
Balzarini
J
J
Hernández
A I
AI
De Clercq
E
E
Priego
E M
EM
Camarasa
M J
MJ
Pérez-Pérez
M J
MJ
D000495:Allosteric Site; D000818:Animals; D002645:Chickens; D002823:Chorion / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004926:Escherichia coli / Q000201:enzymology; D006801:Humans; D020128:Inhibitory Concentration 50; D007288:Inosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D009369:Neoplasms / Q000188:drug therapy; D009389:Neovascularization, Pathologic; D010710:Phosphates; D011743:Pyrimidines / Q000737:chemistry; D013939:Thymidine Phosphorylase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014320:Triphenylmethyl Compounds / Q000494:pharmacology*
C514067:5'-O-tritylinosine; D004791:Enzyme Inhibitors; D010710:Phosphates; D011743:Pyrimidines; D014320:Triphenylmethyl Compounds; D007288:Inosine; D013939:Thymidine Phosphorylase