<p>(<b>A</b>) Hemin-agarose binding assay showing potential of SjCP3842 to bind heme on hemin-agarose beads. (<b>B</b>) Hemin-agarose binding assay confirmed by immunoblotting using three candidates. ‘U’: unbound, ‘W’: last wash, ‘E’: eluates. (<b>C</b>) Identification of SjCP3842 in heme-binding protein fractions from parasite crude extracts (SWA). (<b>D</b>) Estimation of the amount of heme bound using peroxidase activity of bound heme. Standard curve (linear graph) of peroxidase activity of known concentrations of hemin was used to estimate the amount of bound heme. (<b>E</b>) Differential spectral titration of protein-heme interaction using 10 µM of heme and increasing concentrations of the protein (0 to 28 µM). Soret peak was red shifted from 388 nm to 412 nm, and absorption maximum increased with increasing accumulation of protein-heme complex. The inset is the heme-binding curve constructed by plotting ΔA<sub>412</sub> versus protein concentration, showing 1∶1 stoichiometry.</p
