Figure 9.

Disruption of Fasciola nMF6p/FhHDM-1·hemin complexes of high molecular mass by apoMYO. Size-exclusion chromatography analyses were performed on an FPLC system equipped with a Superdex 75 HR 10/30 column and continuous monitoring at 282 (blue line) and 395 nm (red line). Elution volumes are indicated above each peak fraction. A and B, elution profiles of F. hepatica SAs subjected to SEC alone (A) or mixed with an excess of apoMYO (B). After mixing the SAs with apoMYO, most of the heme signal observed at 8.11 ml in A was eluted at 12.07 ml (B). MALDI-TOF MS confirmed the presence of nMF6p/FhHDM-1 in peak II, MYO in peak III, and a mixture of nMF6p/FhHDM-1 and MYO in peak IV. C and D, elution profiles of myoglobin prior to (C) and after (D) acidification and MEK extraction to obtain apoMYO. Most heme signal (red line) disappeared after MEK treatment, and the protein (black line) eluted as a main peak in both chromatograms (elution volume, 12.7 and 11.88 ml, respectively). A small peak (elution volume, 10.2 ml), probably corresponding to apoMYO aggregates, is also observed in D. mAU, milliabsorbance units.