title: FH8--a small EF-hand protein from Fasciola hepatica.
abstract: Vaccine and drug development for fasciolasis rely on a thorough understanding of the mechanisms involved in parasite-host interactions. FH8 is an 8 kDa protein secreted by the parasite Fasciola hepatica in the early stages of infection. Sequence analysis revealed that FH8 has two EF-hand Ca(2+)-binding motifs, and our experimental data show that the protein binds Ca(2+) and that this induces conformational alterations, thus causing it to behave like a sensor protein. Moreover, FH8 displays low affinity for Ca(2+) (K(obs) = 10(4) m(-1)) and is highly stable in its apo and Ca(2+)-loaded states. Homology models were built for FH8 in both states. It has only one globular domain, with two binding sites and appropriate groups in the positions for coordination of the metal ions. However, an unusually high content of positively charged amino acids in one of the binding sites, when compared with the prototypical sensor proteins, potentially affects the protein's affinity for Ca(2+). The only Cys present in FH8, conserved in the homologous proteins of other helminth parasites, is located on the surface, allowing the formation of dimers, detected on SDS gels. These findings reflect specificities of FH8, which are most probably related to its roles both in the parasite and in the host.
authors.affiliation: IBMC, Institute for Molecular and Cell Biology, University of Porto, Portugal.
authors.forename: Hugo
authors.initials: H
authors.lastname: Fraga
authors.forename: Tiago Q
authors.initials: TQ
authors.lastname: Faria
authors.forename: Filipe
authors.initials: F
authors.lastname: Pinto
authors.forename: Agostinho
authors.initials: A
authors.lastname: Almeida
authors.forename: Rui M M
authors.initials: RM
authors.lastname: Brito
authors.forename: Ana M
authors.initials: AM
authors.lastname: Damas
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000378:metabolism; D003001:Cloning, Molecular; D020832:EF Hand Motifs; D004591:Electrophoresis, Polyacrylamide Gel; D005210:Fasciola hepatica / Q000378:metabolism*; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D013050:Spectrometry, Fluorescence
chemical_list: D015801:Helminth Proteins; D002118:Calcium