RESUMO
J-domain proteins (JDPs) are obligate cochaperones of Hsp70s. The Class A JDP Apj1 of the yeast cytosol has an unusually complex region between the N-terminal J-domain and the substrate binding region-often called the Grich or GF region in Class A and B JDPs because of its typical abundance of glycine. The N-terminal 161-residue Apj1 fragment is known to be sufficient for Apj1 function in prion curing, driven by the overexpression of Hsp104. Further analyzing the N-terminal segment of Apj1, we found that a 90-residue fragment that includes the 70-residue J-domain and the adjacent 12-residue glutamine/alanine (Q/A) segment is sufficient for curing. Furthermore, the 121-residue fragment that includes the Grich region was sufficient to not only sustain the growth of cells lacking the essential Class B JDP Sis1 but also enabled the maintenance of several prions normally dependent on Sis1 for propagation. A J-domain from another cytosolic JDP could substitute for the Sis1-related functions but not for Apj1 in prion curing. Together, these results separate the functions of JDPs in prion biology and underscore the diverse functionality of multi-domain cytosolic JDPs in yeast.
RESUMO
AbstractAntioxidants have important physiological roles in limiting the amount of oxidative damage that an organism experiences. One putative antioxidant is biliverdin, a pigment that is most commonly associated with the blue or green colors of avian eggshells. However, despite claims that biliverdin functions as an antioxidant, neither the typical physiological concentrations of biliverdin in most species nor the ability of biliverdin to oppose oxidative damage at these concentrations has been examined. Therefore, we quantified biliverdin in the plasma of six bird species and found that they circulated levels of biliverdin between 0.02 and 0.5 µM. We then used a pool of plasma from northern bobwhite quail (Colinus virginianus) and spiked it with one of seven different concentrations of biliverdin, creating plasma-based solutions ranging from 0.09 to 231 µM biliverdin. We then compared each solution's ability to oppose oxidative damage in response to hydrogen peroxide relative to a control addition of water. We found that hydrogen peroxide consistently induced moderate amounts of oxidative damage (quantified as reactive oxygen metabolites) but that no concentration of biliverdin ameliorated this damage. However, biliverdin and hydrogen peroxide interacted, as the amount of biliverdin in hydrogen peroxide-treated samples was reduced to approximately zero, unless the initial concentration was over 100 µM biliverdin. These preliminary findings based on in vitro work indicate that while biliverdin may have important links to metabolism and immune function, at physiologically relevant concentrations it does not detectably oppose hydrogen peroxide-induced oxidative damage in plasma.
