1. A physicochemical investigation on the metal binding properties of TtSmtB, a thermophilic member of the ArsR/SmtB transcription factor family
- Author
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Emilia Pedone, Immacolata Antonucci, Angela Amoresano, Danila Limauro, Gabriella Fiorentino, Simonetta Bartolucci, Luciano Pirone, Giovanni Gallo, Patrizia Contursi, Gallo, G, Antonucci, I, Pirone, L, Amoresano, A, Contursi, P, Limauro, D, Pedone, E, Bartolucci, S, and Fiorentino, G.
- Subjects
Models, Molecular ,Chemical Phenomena ,Protein Conformation ,Recombinant Fusion Proteins ,Allosteric regulation ,Repressor ,Metal Binding Site ,02 engineering and technology ,Calorimetry ,Biochemistry ,03 medical and health sciences ,Structural Biology ,Amino Acid Sequence ,Molecular Biology ,Transcription factor ,Gene ,Derepression ,030304 developmental biology ,Thermostability ,0303 health sciences ,biology ,Bacteria ,Chemistry ,Spectrum Analysis ,General Medicine ,Thermus thermophilus ,021001 nanoscience & nanotechnology ,biology.organism_classification ,Archaea ,Metals ,Multigene Family ,Thermogravimetry ,Trans-Activators ,0210 nano-technology ,Protein Binding - Abstract
The transcription factors of the ArsR/SmtB family are widespread within the bacterial and archaeal kingdoms. They are transcriptional repressors able to sense a variety of metals and undergo allosteric conformational changes upon metal binding, resulting in derepression of genes involved in detoxification. So far, the molecular determinants of specificity, selectivity, and metal binding mechanism have been scarcely investigated in thermophilic microorganisms. TtSmtB, the only ArsR/SmtB member present in the genome of Thermus thermophilus HB27, was chosen as a model to shed light into such molecular mechanisms at high temperature. In the present study, using a multidisciplinary approach, a structural and functional characterization of the protein was performed focusing on its metal interaction and chemical-physical stability. Our data demonstrate that TtSmtB has two distinct metal binding sites per monomer and interacts with di-tri-penta-valent ions with different affinity. Detailed knowledge at molecular level of protein-metal interaction is remarkable to design metal binding domains as scaffolds in metal-based therapies as well as in metal biorecovery or biosensing in the environment.
- Published
- 2019