The secretory omega-class glutathione transferase OvGST3 from the human pathogenic parasite Onchocerca volvulus.
Onchocerciasis or river blindness, caused by the filarial nematode Onchocerca volvulus, is the second leading cause of blindness due to infectious diseases. The protective role of the omega-class glutathione transferase 3 from O. volvulus (OvGST3) against intracellular and environmental reactive oxygen species has been described previously. In the present study, we continue our investigation of the highly stress-responsive OvGST3. Alternative splicing of two exons and one intron retention generates five different transcript isoforms that possess a spliced leader at their 5'-end, indicating that the mechanism of mature mRNA production involves alternative-, cis- and trans-splicing processes. Interestingly, the first two exons of the ovgst3 gene encode a signal peptide before sequence identity to other omega-class glutathione transferases begins. Only the recombinant expression of the isoform that encodes the longest deduced amino acid sequence (OvGST3/5) was successful, with the purified enzyme displaying modest thiol oxidoreductase activity. Significant IgG1 and IgG4 responses against recombinantly expressed OvGST3/5 were detected in sera from patients with the generalized as well as the chronic hyperreactive form of onchocerciasis, indicating exposure of the secreted protein to the human host's immune system and its immunogenicity. Immunohistological localization studies performed at light and electron microscopy levels support the extracellular localization of the protein. Intensive labeling of the OvGST3 was observed in the egg shell at the morula stage of the embryo, indicating extremely defined, stage-specific expression for a short transient period only.
Institute of Animal Physiology, University of Münster, Münster, Germany. liebaue@uni-muenster.de
Eva
E
Liebau
Jana
J
Höppner
Mareike
M
Mühlmeister
Cora
C
Burmeister
Kai
K
Lüersen
Markus
M
Perbandt
Christel
C
Schmetz
Dietrich
D
Büttner
Norbert
N
Brattig
D000595:Amino Acid Sequence; D000818:Animals; D005091:Exons; D005786:Gene Expression Regulation; D016678:Genome; D005982:Glutathione Transferase / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D008954:Models, Biological; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D017181:Onchocerca volvulus / Q000201:enzymology*; D009855:Onchocerciasis / Q000469:parasitology; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D017386:Sequence Homology, Amino Acid
D011994:Recombinant Proteins; D005982:Glutathione Transferase