CuA-based chimeric T1 copper sites allow for independent modulation of reorganization energy and reduction potential

Attaining rational modulation of thermodynamic and kinetic redox parameters of metalloproteins is a key milestone towards the (re)design of proteins with new or improved redox functions. Here we report that implantation of ligand loops from natural T1 proteins into the scaffold of a CuA protein le...

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Autores principales: Szuster, Jonathan, Zitare, Ulises A., Castro, María A., Leguto, Alcides J., Morgada, Marcos Nicolás, Vila, Alejandro J., Murgida, Daniel H.
Formato: article artículo publishedVersion
Lenguaje:Inglés
Publicado: Royal Society of Chemistry 2021
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Acceso en línea:http://hdl.handle.net/2133/20481
http://hdl.handle.net/2133/20481
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Sumario:Attaining rational modulation of thermodynamic and kinetic redox parameters of metalloproteins is a key milestone towards the (re)design of proteins with new or improved redox functions. Here we report that implantation of ligand loops from natural T1 proteins into the scaffold of a CuA protein leads to a series of distorted T1-like sites that allow for independent modulation of reduction potentials (E°´) and electron transfer reorganization energies (l). On the one hand E°´ values could be fine-tuned over 120 mV without affecting l. On the other, l values could be modulated by more than a factor of two while affecting E°´ only by a few millivolts. These results are in sharp contrast to previous studies that used T1 cupredoxin folds, thus highlighting the importance of the protein scaffold in determining such parameters