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A, loss of ellipticity upon increasing temperature analyzed by CD spectroscopy for PK-WT and the mutant proteins. B, loss of biological activity at 60 °C. C, regain of activity after cooling the denatured proteins on ice. D, activity assay at physiological temperature (37 °C) over a given time period. All assays were done under standard reaction conditions as described under “Experimental Procedures.” mdeg, millidegrees.
