The trematode Fasciola hepatica (Fa. he.) is a common parasite of human and livestock. The hemoglobin (Hb) of Fa. he., a potential immunogen, was chosen for characterization in the search for an effective vaccine. Characterization of trematode Hbs show that they are intracellular single-domain globins with the following remarkable features: (1) Fa. he. expresses two Hb isoforms that differ at two amino acid sites (F1: 119Y/123Q; F2: 119F/123L). Both isoforms are monoacetylated at their N-termini; (2) the genes coding for Fa. he. and Paramphistomum epiclitum (Pa. ep.) Hbs are interrupted by two introns at the conserved positions B12.2 and G7.0.; (3) UV/VIS and resonance Raman spectroscopy identify the recombinant Fa. he. HbF2 as a pentacoordinated high-spin ferrous Hb; (4) electron paramagnetic resonance spectroscopy of cyano-met Fa. he. HbF2 proves that the endogenously bound imidazole has no imidazolate character; (5) the major structural determinants of the globin fold are present, they contain a TyrB10/TyrE7 residue pair on the distal side. Although such distal-site pair is a signature for high oxygen affinity, as shown for Pa. ep. Hb, the oxygen-binding rate parameters for Fa. he. Hb are intermediate between those of myoglobin and those of other trematode Hbs; (6) the three-dimensional structure of recombinant Fa. he. HbF2 from this study closely resembles the three-dimensional structure of Pa. ep. determined earlier. The set of distal-site polar interactions observed in Pa. ep. Hb is matched with small but significant structural adjustments; (7) despite the potential immunogenic character of the fluke Hb, vaccination of calves with recombinant Fa. he. HbF2 failed to promote protection against parasitic infection.
|Titolo:||The hemoglobins of the trematodes Fasciola hepatica and Paramphistomum epiclitum: a molecular biological, physico-chemical, kinetic, and vaccination study|
BOLOGNESI, MARTINO (Penultimo)
|Parole Chiave:||3D structure; Fluke; Hemoglobins; Ligand binding; Spectroscopy; Vaccination|
|Settore Scientifico Disciplinare:||Settore BIO/10 - Biochimica|
|Data di pubblicazione:||2008|
|Digital Object Identifier (DOI):||10.1110/ps.036558.108|
|Appare nelle tipologie:||01 - Articolo su periodico|