Abstract
Heat denaturation of methionine aminopeptidase from a hyperthermophile Pyrococcus furiosus (PfMAP) was studied by differential scanning calorimetry at acid pH. Analysis of the calorimetric data has shown that denaturation of PfMAP is non-equilibrium at heating rates from 0.125 to 2 K min–1. This means that the protein structure at these conditions is metastable and its stability (the apparent temperature of denaturation T m) is under kinetic control. It was shown that heat denaturation of this protein is a one-step kinetic process. The enthalpy of the process and its activation energy were measured as functions of temperature. The obtained data allowed us to estimate the heat capacity increment and the change in the number of bound protons during activation of the molecule. The data also suggest that the conformation of PfMAP at the transition state only slightly differs from its native conformation with respect to compactness, hydration extent and hydroxyl protonation.
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Potekhin, S.A., Ogasahara, K. & Yutani, K. Transition State of Heat Denaturation of Methionine Aminopeptidase from a Hyperthermophile. Journal of Thermal Analysis and Calorimetry 62, 111–122 (2000). https://doi.org/10.1023/A:1010114828690
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DOI: https://doi.org/10.1023/A:1010114828690