Characterization and differential expression of a ferritin protein from Fasciola hepatica

Abstract

Ferritins are proteins that play a central role in maintaining intracellular iron balance. A cDNA clone of Fasciola hepatica (687 bp long) encoding a putative 228-amino acid polypeptide (FhFtn-1) homologous with ferritins of vertebrates and invertebrates was identified. FhFtn-1 contains a conserved motif of the ferroxidase center typical of vertebrate ferritins. Phylogenetic tree analysis showed that FhFtn-1 clusters with two ferritins of Paragonimus westermani, which suggests a common ancestry for the ferritins of these two trematodes. Recombinant FhFtn-1 protein expressed and purified from an Escherichia coli system showed iron-uptake ability. Moreover, FhFtn-1 showed strong reactivity with sera from rabbits infected with F. hepatica for 2–12 weeks, which suggests that this protein could be a potential antigen for immunodiagnosis of fascioliasis. qPCR analysis demonstrated that FhFtn-1-mRNA is expressed at significantly higher levels in adults and unembryonated eggs than in juveniles or miracidia. These results represent the first characterization of a ferritin protein from the liver fluke F. hepatica.

Development of Two Antibody Detection Enzyme-Linked Immunosorbent Assays for Serodiagnosis of Human Chronic Fascioliasis

“…This finding supports our previous studies using experimentally infected rabbits, in which antibodies were detected between 2 to 4 weeks after infection using recombinant FhFtn-1 and FhTP16.5 proteins (29,30). Unfortunately, in the present study we were not able to obtain serum a Sensitivity ϭ 96.6% (95% CI, 88.5% to 99.1%), calculated as true-positive number/ (true-positive number ϩ false-negative number) ϫ 100; specificity ϭ 95.7% (95% CI, 89.5% to 98.3%), calculated as true-negative number/(true-negative number ϩ falsepositive number) ϫ 100.…”

“…HQ316639.1) was cloned into the pRSET A plasmid (Invitrogen, Carlsbad, CA) and used to transform Escherichia coli BL21(DE3) cells (Stratagene, Santa Clara, CA). The recombinant polypeptide was overexpressed as a fusion protein with a His tag and was purified using a HisTrap FF crude affinity column (GE Healthcare Biosciences, Pittsburgh, PA), as described previously (30). cDNA encoding FhTP16.5 (GenBank accession no.…”

“…One of these antigens is a 16.5-kDa tegument-associated protein of unknown function, termed FhTP16.5 (29), and the other is a protein with ferroxidase activity classified as a member of the F. hepatica ferritin protein family (FhFtn-1) (30). Both molecules are differentially expressed during parasite development and have been shown to be highly reactive with sera from experimental animals with acute or chronic infections.…”

Revelation of endogenously bound Fe2+ ions in the crystal structure of ferritin from Escherichia coli

“…The L chain acts as an iron nucleation site and capable of mineralizing iron faster than the heavy chain (H) of ferritin. L ferritin stands out to be higher stability than H ferritin even under acidic and reducing conditions [13,14].…”

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“…Till so far, there are lots of literatures demonstrate that the ferritin protein can offer valid protection mechanisms against the harmful effects of dioxygen damages (Caban-Hernandez et al, 2012), bacterial infections (Rebl et al, 2012) and DNA damages by the UV (Shimmura et al, 1996). It also offers immunity protections for the vertebrates and invertebrates (Ong et al, 2006), and protects the cells from injury under cold stress (0℃ or 4℃) (Zieger and Gupta, 2009).…”