Iron acquisition and virulence in Helicobacter pylori: a major role for FeoB, a high-affinity ferrous iron transporter

The genome sequence of Helicobacter pylori suggests that this bacterium pos sesses several Fe acquisition systems, including both Fe2+- and Fe3+-citrat e transporters. The role of these transporters was investigated by generati ng insertion mutants in feoB, tonB, fecA1 and fecDE. Fe transport in the fe oB mutant was approximate to 10-fold lower than in the wild type (with 0.5 mu M Fe), irrespective of whether Fe was supplied in the Fe2+ or Fe3+ form. In contrast, transport rates were unaffected by the other mutations. Compl ementation of the feoB mutation fully restored both Fe2+ and Fe3+ transport . The growth inhibition exhibited by the feoB mutant in Fe-deficient media was relieved by human holo-transferrin, holo-lactoferrin and Fe3+-dicitrate , but not by FeSO4. The feoB mutant had less cellular Fe and was more sensi tive to growth inhibition by transition metals in comparison with the wild type. Biphasic kinetics of Fe2+ transport in the wild type suggested the pr esence of high- and low-affinity uptake systems. The high-affinity system ( apparent K-s = 0.54 mu M) is absent in the feoB mutant. Transport via FeoB is highly specific for Fe2+ and was inhibited by FCCP, DCCD and vanadate, i ndicating an active process energized by ATP. Ferrozine inhibition of Fe2and Fe3+ uptake implied the concerted involvement of both an Fe3+ reductase and FeoB in the uptake of Fe supplied as Fe3+. Taken together, the results are consistent with FeoB-mediated Fe2+ uptake being a major pathway for H. pylori Fe acquisition. feoB mutants were unable to colonize the gastric mu cosa of mice, indicating that FeoB makes an important contribution to Fe ac quisition by H. pylori in the low-pH, low-O-2 environment of the stomach.