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We expected larger binding isotope effects with AAT than alanine racemase since AAT fully protonates the pyridine nitrogen, whereas in alanine racemase the interaction of the pyridine nitrogen with Arg219 precludes its protonation.[80] Instead of extracting binding isotope effects from a larger analysis of the enzymatic reaction as was done with alanine racemase, we sought to isolate the binding step to enable direct measurement

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