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CD spectroscopy was used to estimate the secondary structures of individual mutants. The CD signal in millidegrees was converted to mean-residue ellipticity [θ]. Far-UV CD spectra of (A) the hydrophobic core double mutants at pH4.0, and (B) at pH 6.0. (C) The salt bridge mutants at pH 4.0, and (D) at pH 6.0.
