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Using the measured pKa value for the 2′-hydroxy group and the pKa of hydronium ion as an estimate for the pKa of the 2′-oxonium ion, we estimate βnuc as 0.59. This value likely underestimates βnuc for two reasons: 1) the neutral hydroxy group would experience less electrostatic repulsion with the reaction center than the oxyanion; 2 the nucleotidyl context of the hydroxy group likely decreases the pKa value of the 2′-oxonium ion relative to a hydronium ion. For example, ammonium ion has a pKa value of 10, but the 2′-nucleotidyl ammonium ion ionizes with pKa ≈ 6.2.[18
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[18]
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