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• W2032766238 endingPage "350" @default.
• W2032766238 startingPage "342" @default.
• W2032766238 abstract "Thymidylate synthase has been purified greater than 4000-fold from a human colon adenocarcinoma maintained as a xenograft in immune-deprived mice. In this disease, the enzyme is an important target for the cytotoxic action of 5-fluorouracil, which is influenced by the reduced folate substrate CH2-H4PteGlu. Due to the importance of this interaction, and the existence in cells of folate species as polyglutamyl forms, the interaction of folylpolyglutamates with thymidylate synthase was examined. Polyglutamates of PteGlu were used as inhibitors, and the interaction of CH2-H4PteGlu polyglutamates as substrates or in an inhibitory ternary complex were also examined. Using PteGlu1-7, Ki values were determined. A maximal 125-fold decrease in Ki was observed between PteGlu1 and PteGlu4; further addition of up to three glutamyl residues did not result in an additional decrease in Ki. Despite the increased binding affinity of folypolyglutamates for this enzyme, no change in the Km values for either dUMP (3.6 microM) or CH2-H4PteGlu (4.3 microM) were detected when polyglutamates of [6R]CH2-H4PteGlu were used as substrates. Product inhibition studies demonstrated competitive inhibition between dTMP and dUMP in the presence of CH2-H4PteGlu5. In addition, CH2-H4PteGlu4 stabilized an inhibitory ternary complex formed between FdUMP, thymidylate synthase, and CH2-H4PteGlu4. Thus the data do not support a change in the order of substrate binding and product release upon polyglutamylation of CH2-H4PteGlu reported for non-human mammalian enzyme. This is the first study to characterize kinetically thymidylate synthase from a human colon adenocarcinoma." @default.
• W2032766238 created "2016-06-24" @default.
• W2032766238 creator A5022043593 @default.
• W2032766238 creator A5031291620 @default.
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• W2032766238 date "1988-01-01" @default.
• W2032766238 modified "2023-10-16" @default.
• W2032766238 title "Characteristics of thymidylate synthase purified from a human colon adenocarcinoma" @default.
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• W2032766238 doi "https://doi.org/10.1016/0003-9861(88)90458-4" @default.
• W2032766238 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/3341747" @default.
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