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Holo-hCA II rotameric states of His64: (A) 0 s, (B) 50 s, (C) 3 min, (D) 10 min, (E) 25 min, and (F) 1 h. Electron density 2Fo-Fc (blue) and Fo-Fc (green) maps are contoured at 1.5σ and 3.0σ, respectively. The 2Fo-Fc maps for waters are highlighted in pink for clarity. On CO2 release and gradual dissipation of W2′, the His64 side chain shifts from the out to the in position. Solvent and amino acids are as labeled.
