FIGURE 4.
Heme transport analysis of wild-type and mutant CeHRG-1. A, the hem1Δ(6D) strain was transformed with empty vector pYes-DEST52, CeHRG-1-HA, and its mutants. A spot growth assay were performed with the indicated conditions. Plates were incubated at 30 °C for 3 days prior to imaging. B, the hem1Δfre1Δfre2ΔMET3-FRE1 strain was transformed with empty vector pYes-DEST52, CeHRG-1, and its mutants. A ferric reductase assay was performed with the indicated concentrations of hemin. Ferric reductase activity (nanomoles/106cells/min) was measured and normalized to the activity of vector. C, the hem1Δ(6D) strain co-transformed with pCYC1-LacZ and empty vector pYes-DEST52, CeHRG-1, or its mutants. β-Galactosidase activity was measured. Error bars represent the ±S.E. from three independent experiments. *, p < 0.05; **, p < 0.01; and ***, p < 0.001.