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pD profile of pyruvate C3 proton exchange reaction in HpaI aldolase. Assays were performed in deuterated constant ionic strength buffer containing 100 mm Tris, 50 mm acetic acid, 50 mm MES, and 1.0 mm CoCl2 with pD ranging from 5.4 to 9.4. The data were fitted to Equation 1 by non-linear regression using Leonora and showed a pK1 of 6.5 ± 0.1.
