Korean J Parasitol.  2018 Feb;56(1):81-86. 10.3347/kjp.2018.56.1.81.

Comparative Characterization of Four Calcium-Binding EF Hand Proteins from Opisthorchis viverrini

• Emmanoch P
• Kosa N
• Vichasri-Grams S
• Tesana S
• Grams R
• Geadkaew-Krenc A

• KMID: 2408529
• DOI: http://doi.org/10.3347/kjp.2018.56.1.81

Abstract

Four isoforms of calcium binding proteins containing 2 EF hand motifs and a dynein light chain-like domain in the human liver fluke Opisthorchis viverrini, namely OvCaBP1, 2, 3, and 4, were characterized. They had molecular weights of 22.7, 21.6, 23.7, and 22.5 kDa, respectively and showed 37.2-42.1% sequence identity to CaBP22.8 of O. viverrini. All were detected in 2- and 4-week-old immature and mature parasites. Additionally, OvCaBP4 was found in newly excysted juveniles. Polyclonal antibodies against each isoform were generated to detect the native proteins in parasite extracts by Western blot analysis. All OvCaBPs were detected in soluble and insoluble crude worm extracts and in the excretory-secretory product, at approximate sizes of 21-23 kDa. The ion-binding properties of the proteins were analyzed by mobility shift assays with the divalent cations Ca²âº, Mg²âº, Zn²âº, and Cu²+. All OvCaBPs showed mobility shifts with Ca²âº and Zn²âº. OvCaBP1 showed also positive results with Mg²âº and Cu²âº. As tegumental proteins, OvCaBP1, 2, and 3 are interesting drug targets for the treatment of opisthorchiasis.

Keyword

Opisthorchis viverrini; Platyhelminthes; EF hand motif; calcium-binding; dynein light chain; tegument

MeSH Terms

Antibodies
Blotting, Western
Calcium-Binding Proteins
Cations, Divalent
Dyneins
EF Hand Motifs*
Electrophoretic Mobility Shift Assay
Fasciola hepatica
Humans
Molecular Weight
Opisthorchiasis
Opisthorchis*
Parasites
Protein Isoforms
Antibodies
Calcium-Binding Proteins
Cations, Divalent
Dyneins
Protein Isoforms