Cooperation between two periplasmic copper chaperones is required for full activity of the cbb 3 -type cytochrome c oxidase and copper homeostasis in Rhodobacter capsulatus
Copper (Cu) is an essential micronutrient that functions as a cofactor in several important enzymes, like respiratory heme-copper oxygen reductases. Yet, Cu is also toxic and therefore cells engage a highly coordinated Cu uptake and delivery system to prevent the accumulation of toxic Cu concentrations. In the current work we analyzed Cu delivery to the cbb 3 -type cytochrome c oxidase ( cbb 3 -Cox) of Rhodobacter capsulatus . We identified the PCu A C-like periplasmic chaperone PccA and analyzed its contribution to cbb 3 -Cox assembly. Our data demonstrate that PccA is a Cu-binding protein with a preference for Cu(I), which is required for efficient cbb 3 -Cox assembly, in particular at low Cu concentrations. By using in vivo and in vitro crosslinking we show that PccA forms a complex with the Sco1-homologue SenC. This complex is stabilized in the absence of the cbb 3 -Cox specific assembly factors CcoGHIS. In cells lacking SenC, the cytoplasmic Cu content is significantly increased, but the simultaneous absence of PccA prevents this Cu accumulation. Lastly, these data demonstrate that the interplay between PccA and SenC is not only required for Cu delivery during cbb 3 -Cox assembly, but that it also regulates Cu homeostasis in R. capsulatus .