Alcohol dehydrogenase from the hyperthermophilic archaeon Pyrobaculum aerophilum: Stability at high temperature

作者：Ausili Alessio^*; Vitale Annalisa; Labella Tullio; Rosso Francesco; Barbarisi Alfonso; Gomez Fernandez Juan C; D' Auria Sabato

来源：Archives of Biochemistry and Biophysics, 2012, 525(1): 40-46.

DOI：10.1016/j.abb.2012.05.019

• FTIR
• DSC
• Thermostable proteins
• ADH
• Stability

The structural and stability properties of a novel zinc-dependent alcohol dehydrogenase from the hyperthermophilic archaeon Pyrobaculum aerophilum (PyAeADHII) were investigated by Fourier transformed infrared spectroscopy (FTIR). This enzyme is a thermostable homo-tetramer belonging to the GroES-fold motif proteins characterized by an irregular beta-barrel conformation. Our results revealed a protein with a secondary structure rich in beta-sheet (32% of the total secondary elements) and it showed a three-step thermal unfolding pathway. The complete enzyme denaturation was preceded by the formation of a relaxed tertiary/quaternary structure and previously by an excited native-like conformation. Two-dimensional correlation spectroscopy analysis (2D-COS) and differential scanning calorimetry (DSC) experiments supported these data and allowed us to determine the protein melting temperature at 96.9 degrees C as well as to suggest the sequence of the events that occurred during the protein denaturation process.

• 出版日期2012-9-1