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. Author manuscript; available in PMC: 2021 Sep 15.
Published in final edited form as: Biochem J. 2020 Feb 14;477(3):727–745. doi: 10.1042/BCJ20190874

Figure 8. Thermal stability of mature, purified TPP1.

Figure 8.

(A) Untagged WT TPP1 CD spectra. Wavelength scans from 210-260 nm were performed on mature TPP1 at increasing temperatures and plotted against ellipticity. (B) Thermal denaturation of WT TPP1 determined by CD and enzymatic activity. Thermal denaturation experiments were carried out by monitoring the ellipticity of mature native TPP1 at 222 nm as the temperature was increased from 4°C to 90°C in 2°C increments. An equivalent sample of mature WT TPP1 was also heated from 40°C to 90°C in 5°C increments with TPP1 activity measured at each increment. (C) Change in melting temperature of his-tagged TPP1 variants compared to untagged WT TPP1. CD and enzyme activity assays where carried out for each TPP1 variant in the same manner as outlined in (B). Error bars display standard deviation in all cases. All thermal denaturation experiments were performed in triplicate.