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Average MD structures of the (a) ES and (b) ES′ states in AHA, with the key interaction between Asp264 and the substrate hydroxyls present and absent, respectively. (c) Calculated probability density7 of the Asp264 Oδ2–substrate O2 distance as a function of temperature. (d) Calculated probability density for the same interaction (Asp300–substrate) in the mesophilic PPA ortholog, where it breaks at higher temperatures.
