TY - JOUR
T1 - Interactions of 2′-fluoro-substituted dUMP analogues with thymidylate synthase
AU - Ziemkowski, Przemysław
AU - Felczak, Krzysztof
AU - Poznański, Jarosław
AU - Kulikowski, Tadeusz
AU - Zieliński, Zbigniew
AU - Cieśla, Joanna
AU - Rode, Wojciech
PY - 2007/10/12
Y1 - 2007/10/12
N2 - A series of 2′-fluoro-substituted dUMP/FdUMP analogues were synthesized, their interaction with human recombinant thymidylate synthase investigated, and structural 1H and 19F NMR study of the corresponding nucleosides performed. While 2′-F-dUMP (fluorine in the "down" configuration), in striking contrast to 2′-F-ara-UMP (fluorine in the "up" configuration) and 2′,2′′-diF-dUMP, showed substrate activity, 2′-F-ara-UMP and 2′,2′′-diF-dUMP were classic inhibitors, and 2′,5-diF-ara-UMP behaved as a strong slow-binding inhibitor, suggesting the 2′-F substituent in the "up" position to interfere with the active center cysteine thiol addition to the pyrimidine C(6) and the pyrimidine C(5)-F to prevent this interference. In support, the direct through space heteronuclear coupling JHF was observed for the fluorine "up" derivatives, 2′-F-ara-U and 2′,5-diF-ara-U, causing the splitting of the H(6) resonance lines. The absence of such splitting in 2′,2′′-diF-dUrd, indicating an unusual orientation of the base in relation to the furanose, was associated with an exceptionally weak interaction with the enzyme.
AB - A series of 2′-fluoro-substituted dUMP/FdUMP analogues were synthesized, their interaction with human recombinant thymidylate synthase investigated, and structural 1H and 19F NMR study of the corresponding nucleosides performed. While 2′-F-dUMP (fluorine in the "down" configuration), in striking contrast to 2′-F-ara-UMP (fluorine in the "up" configuration) and 2′,2′′-diF-dUMP, showed substrate activity, 2′-F-ara-UMP and 2′,2′′-diF-dUMP were classic inhibitors, and 2′,5-diF-ara-UMP behaved as a strong slow-binding inhibitor, suggesting the 2′-F substituent in the "up" position to interfere with the active center cysteine thiol addition to the pyrimidine C(6) and the pyrimidine C(5)-F to prevent this interference. In support, the direct through space heteronuclear coupling JHF was observed for the fluorine "up" derivatives, 2′-F-ara-U and 2′,5-diF-ara-U, causing the splitting of the H(6) resonance lines. The absence of such splitting in 2′,2′′-diF-dUrd, indicating an unusual orientation of the base in relation to the furanose, was associated with an exceptionally weak interaction with the enzyme.
KW - 2′-Fluoro-substituted dUMP analogues
KW - NMR
KW - Thymidylate synthase
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U2 - 10.1016/j.bbrc.2007.07.097
DO - 10.1016/j.bbrc.2007.07.097
M3 - Article
C2 - 17692822
AN - SCOPUS:34548086001
SN - 0006-291X
VL - 362
SP - 37
EP - 43
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -