Browsing by Author "Burgos-Bravo, Francesca"

Now showing 1 - 2 of 2
  • No Thumbnail Available
    Item
    Effect of temperature and nucleotide on the binding of BiP chaperone to a protein substrate
    (2023) Rivera, Maira; Burgos-Bravo, Francesca; Engelberger, Felipe; Asor, Roi; Lagos-Espinoza, Miguel I. A.; Figueroa, Maximiliano; Kukura, Philipp; Ramirez-Sarmiento, Cesar A.; Baez, Mauricio; Smith, Steven B.; Wilson, Christian A. M.
    BiP (immunoglobulin heavy-chain binding protein) is a Hsp70 monomeric ATPase motor that plays broad and crucial roles in maintaining proteostasis inside the cell. Structurally, BiP is formed by two domains, a nucleotide-binding domain (NBD) with ATPase activity connected by a flexible hydrophobic linker to the substrate-binding domain. While the ATPase and substrate binding activities of BiP are allosterically coupled, the latter is also dependent on nucleotide binding. Recent structural studies have provided new insights into BiP's allostery; however, the influence of temperature on the coupling between substrate and nucleotide binding to BiP remains unexplored. Here, we study BiP's binding to its substrate at the single molecule level using thermo-regulated optical tweezers which allows us to mechanically unfold the client protein and explore the effect of temperature and different nucleotides on BiP binding. Our results confirm that the affinity of BiP for its protein substrate relies on nucleotide binding, by mainly regulating the binding kinetics between BiP and its substrate. Interestingly, our findings also showed that the apparent affinity of BiP for its protein substrate in the presence of nucleotides remains invariable over a wide range of temperatures, suggesting that BiP may interact with its client proteins with similar affinities even when the temperature is not optimal. Thus, BiP could play a role as a "thermal buffer" in proteostasis.
  • Thumbnail Image
    Item
    Generation of superoxide radicals by copper-glutathione complexes: Redox-consequences associated with their interaction with reduced glutathione
    (2009) Speisky, Hernán; Gomez, Maritza; Burgos-Bravo, Francesca; López Alarcón, Camilo Ignacio; Jullian, Carolina; Olea-Azar, Claudio; Aliaga Miranda, Margarita Elly