Structural characterization of the human DjC20/HscB cochaperone in solution.

J-domain proteins (JDPs) form a very large molecular chaperone family involved in proteostasis processes, such as protein folding, trafficking through membranes and degradation/disaggregation. JDPs are Hsp70 co-chaperones capable of stimulating ATPase activity as well as selecting and presenting client proteins to Hsp70. In mitochondria, human DjC20/HscB (a type III JDP that possesses only the conserved J-domain in some region of the protein) is involved in [FeS] protein biogenesis and assists human mitochondrial Hsp70 (HSPA9). Human DjC20 possesses a zinc-finger domain in its N-terminus, which closely contacts the J-domain and appears to be essential for its function. Here, we investigated the hDjC20 structure in solution as well as the importance of Zn+2 for its stability. The recombinant hDjC20 was pure, folded and capable of stimulating HSPA9 ATPase activity. It behaved as a slightly elongated monomer, as attested by small-angle X-ray scattering and SEC-MALS. The presence of Zn2+ in the hDjC20 samples was verified, a stoichiometry of 1:1 was observed, and its removal by high concentrations of EDTA and DTPA was unfeasible. However, thermal and chemical denaturation in the presence of EDTA led to a reduction in protein stability, suggesting a synergistic action between the chelating agent and denaturators that facilitate protein unfolding depending on metal removal. These data suggest that the affinity of Zn+2 for the protein is very high, evidencing its importance for the hDjC20 structure.

Solubility of Cd, Cr, Cu, Ni, and Pb and Its Correlation with Total Polyphenols and Soluble Melanoidins in Hot Infusions of Green and Roasted Mate.

The total and soluble contents of three potentially toxic elements (PTEs) (Cd, Cr, and Pb) and two essential elements (Cu and Ni) in commercial green and roasted mate (Ilex paraguariensis St. Hil.) and their hot infusions were accessed by inductively coupled plasma mass spectrometry. Total polyphenolic (TP) contents in the infusions were accessed by Folin-Ciocalteu method, and the presence of soluble melanoidins (SM) was evaluated by measuring absorbances at 420 nm. The soluble element contents in hot infusions of green mate were present in the following increasing order: Pb < Cd < Cr < Cu, and Ni. For hot infusions of roasted mate, the increasing order was: Cu < Pb < Cd < Cr < Ni. Except for Cr, hot infusions of roasted mate supply lower contents of the soluble PTEs than hot infusions of green mate. Cadmium was the only of evaluated trace element found in hot infusions of green and roasted mate as well as in some of their mate samples above the maximum allowed contents established by Brazilian legislation. Despite this, polyphenol and melanoidin contents in green and roasted mate infusions may reduce Cd bioavailability. The soluble trace elements that correlated directly with total polyphenols in hot infusions of green or roasted mate are indirectly correlated with the presence of soluble melanoidins. Moreover, this is the first study to evaluate the correlations of soluble forms of Cd, Cr, Cu, Ni, and Pb with the presence of SM and TP contents in hot infusions of green and roasted mate.