Interaction of <i>Pf</i>ATM1 with ISC and CIA proteins.

<p>(<b>A</b>) <i>Pf</i>IscU and <i>Pf</i>IscA2 after thrombin cleavage for removal of GST from the purified fusion proteins. (<b>B-C</b>) Size exclusion chromatography of partially purified <i>Pf</i>NBP35 (B) and <i>Pf</i>CFD1 (C) through S75 column. Absorbance at 280 nm is plotted against elution volume. Inset, Molecular weight standard plot for S75. (<b>D</b>) Coomassie-stained SDS-PAGE of purified recombinant <i>Pf</i>NBP35 (~40 kDa) and (<b>E</b>) Purified recombinant <i>Pf</i>CFD1 (~57.5 kDa). (<b>F</b>) 6–10% native PAGE to detect complexation of <i>Pf</i>NBP35 and <i>Pf</i>CFD1. (<b>G</b>) 6–8% native PAGE to detect in vitro complexation of <i>Pf</i>ATM1 dimer (lane 1) with <i>Pf</i>CFD1 (lane 2) and <i>Pf</i>NBP35 (lane 3). The <i>Pf</i>NBP35-<i>Pf</i>CFD1 heterotetramer (lane 5) complexed with the <i>Pf</i>ATM1 dimer (lane 4). (<b>H)</b> 10–12% native PAGE to detect in vitro complexation of <i>Pf</i>ATM1-NTE and <i>Pf</i>ATM1-CTD with <i>Pf</i>NBP35. (<b>I</b>) Anti-<i>Pf</i>NBP35 serum detects a specific band of the expected size of ~51 kDa in the parasite lysate. (<b>J</b>) Pull-down from <i>P</i>. <i>falciparum</i> lysate using <i>Pf</i>ATM1-NTE as bait. Anti-6XHis antibodies and anti-<i>Pf</i>NBP35 serum were used to detect <i>Pf</i>ATM1-NTE and <i>Pf</i>NBP35, respectively in western blots. (<b>K</b>) <i>Ec</i>EngA used as negative control bait for the pull-down experiment in (<b>J</b>).</p> <p>(PDF)</p>