Alcohol dehydrogenase from the hyperthermophilic archaeon Pyrobaculum aerophilum: stability at high temperature |
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Authors: | Ausili Alessio Vitale Annalisa Labella Tullio Rosso Francesco Barbarisi Alfonso Gómez-Fernández Juan C D'Auria Sabato |
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Affiliation: | Laboratory for Molecular Sensing, IBP-CNR, Naples, Italy. a.ausili@ibp.cnr.it |
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Abstract: | 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 β-barrel conformation. Our results revealed a protein with a secondary structure rich in β-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 °C as well as to suggest the sequence of the events that occurred during the protein denaturation process. |
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