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Cofactor analog binding to the C146S-dUMP complex by ITC. Raltitrexed was titrated into the pre-formed nucleotide complex at 25 °C (Top), 15 °C (Middle), and 5 °C (Bottom). The inset histograms plot the cooperativity parameter, ρ, which is a ratio of the two intrinsic association constants for binding to the free and singly bound enzyme.
