Iron transport and its relation to heme biosynthesis in Rhodopseudomonas sphaeroides

Abstract

The uptake of iron supplied as ferric citrate or ferric parabactin was examined in aerobically grown whole cells and vesicles of Rhodopseudomonas sphaeroides. Inner and outer membrane fractions from R. sphaeroides contained no membrane proteins which were inducible by growth in low-iron medium. Vesicles composed of the inner membrane and devoid of outer membrane and periplasmic proteins were able to transport iron supplied as ferric citrate and ferric parabactin. This uptake required the presence of NADH. When the electrical component of the proton motive force was depleted in whole cells, the uptake of iron supplied as ferric parabactin was completely inhibited. The uptake of iron supplied as ferric citrate was inhibited by gallium citrate; however, Ga3+ was not transported. The relationship between iron uptake and heme synthesis was examined by treating whole cells with N-methylprotoporphyrin which inhibits ferrochelatase, the enzyme which inserts ferrous iron into protoporphyrin to form heme. This treatment reduced ferrochelatase activity by 82% but had no effect on iron uptake, indicating that iron uptake and heme synthesis are not directly coupled. The fate of transported iron was investigated by measuring intracellular concentrations of heme and nonheme iron. It was determined that newly transported iron exists primarily as nonheme iron.