Urea and thermal denaturation experiments were performed on a JASCO J-1500 circular dichroism (CD) spectrophotometer, and the data were fitted using the program KaleidaGraph. All lyophilized protein samples were dissolved in a buffer containing 10 mM sodium phosphate (pH 6.8). For each complex, 40 μM of the CID domain was mixed with 15 μM NCBD domain. The temperature was set to 298 K for the urea denaturation experiments. The temperature was varied between 278 and 363 K for the thermal denaturation. The ellipticity at 222 nm was monitored for both measurements. The data were then fitted to the standard equations describing two-state unfolding by chemical denaturant and temperature, respectively (17).

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