1. Substitution of the Native Zn(II) with Cd(II), Co(II) and Ni(II) Changes the Downhill Unfolding Mechanism of Ros87 to a Completely Different Scenario
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Sara García-Viñuales, Carla Isernia, Roberto Fattorusso, Sabrina Esposito, Laura Zaccaro, Rinaldo Grazioso, Danilo Milardi, Rosa Iacovino, Luigi Russo, Gianluca D'Abrosca, Gaetano Malgieri, Grazioso, R., Garcia-Vinuales, S., Russo, L., D'Abrosca, G., Esposito, S., Zaccaro, L., Iacovino, R., Milardi, D., Fattorusso, R., Malgieri, G., and Isernia, C. more...
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Models, Molecular ,0301 basic medicine ,Protein Folding ,Magnetic Resonance Spectroscopy ,folding mechanism ,01 natural sciences ,lcsh:Chemistry ,lcsh:QH301-705.5 ,Spectroscopy ,Zinc finger ,Chemistry ,Zinc Fingers ,General Medicine ,cobalt ,Computer Science Applications ,DNA-Binding Proteins ,Folding (chemistry) ,Zinc ,visual_art ,visual_art.visual_art_medium ,Thermodynamics ,Downhill folding ,cadmium ,Metal ions in aqueous solution ,chemistry.chemical_element ,010402 general chemistry ,Article ,Catalysis ,Ion ,Inorganic Chemistry ,Metal ,03 medical and health sciences ,nickel ,Bacterial Proteins ,Amino Acid Sequence ,Physical and Theoretical Chemistry ,Molecular Biology ,Binding Sites ,zinc finger ,Organic Chemistry ,DNA-binding domain ,0104 chemical sciences ,Repressor Proteins ,body regions ,Crystallography ,030104 developmental biology ,lcsh:Biology (General) ,lcsh:QD1-999 ,Agrobacterium tumefaciens ,Spectrophotometry, Ultraviolet - Abstract
The structural effects of zinc replacement by xenobiotic metal ions have been widely studied in several eukaryotic and prokaryotic zinc-finger-containing proteins. The prokaryotic zinc finger, that presents a bigger &beta, &beta, &alpha, domain with a larger hydrophobic core with respect to its eukaryotic counterpart, represents a valuable model protein to study metal ion interaction with metallo-proteins. Several studies have been conducted on Ros87, the DNA binding domain of the prokaryotic zinc finger Ros, and have demonstrated that the domain appears to structurally tolerate Ni(II), albeit with important structural perturbations, but not Pb(II) and Hg(II), and it is in vitro functional when the zinc ion is replaced by Cd(II). We have previously shown that Ros87 unfolding is a two-step process in which a zinc binding intermediate converts to the native structure thorough a delicate downhill folding transition. Here, we explore the folding/unfolding behaviour of Ros87 coordinated to Co(II), Ni(II) or Cd(II), by UV-Vis, CD, DSC and NMR techniques. Interestingly, we show how the substitution of the native metal ion results in complete different folding scenarios. We found a two-state unfolding mechanism for Cd-Ros87 whose metal affinity Kd is comparable to the one obtained for the native Zn-Ros87, and a more complex mechanism for Co-Ros87 and Ni-Ros87, that show higher Kd values. Our data outline the complex cross-correlation between the protein&ndash, metal ion equilibrium and the folding mechanism proposing such an interplay as a key factor in the proper metal ion selection by a specific metallo-protein. more...
- Published
- 2020
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