The peroxidoxin 2 protein of the human parasite Onchocerca volvulus: recombinant expression, immunolocalization, and demonstration of homologous molecules in other species.
The peroxidoxin protein of the filarial parasite Onchocerca volvulus (OvPXN-2) belongs to a group of highly conserved antioxidant molecules. For a more detailed characterization of this protein and for determination of its expression pattern the OvPXN-2 protein was recombinantly expressed as a His-tagged protein. Under reducing conditions the recombinant protein had an apparent molecular mass of 28 kDa. Considering the size of the His-tag and the FLAG epitope introduced to the recombinant protein, this size is in agreement with that of the native protein identified in O. volvulus extract. Antiserum raised against the recombinant protein was used for immunolocalization. In O. volvulus the antigen is predominantly expressed in the hypodermis and particularly the lateral and median chords show high levels of expression. The protein is also expressed strongly in the hypodermis of infective larvae and more weakly in microfilariae. Related cross-reacting proteins were detected in several Onchocerca species and other filariae. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis in combination with Western blotting revealed proteins with almost identical mobility in extracts prepared from O. ochengi, O. gibsoni, and Dirofilaria immitis.
Zipfel
P F
PF
Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany. zipfel@uke.uni-hamburg.de
Schrum
S
S
Bialonski
A
A
Büttner
D W
DW
D000818:Animals; D015139:Blotting, Southern; D015153:Blotting, Western; D003429:Cross Reactions; D004183:Dirofilaria immitis / Q000737:chemistry; D017238:Genes, Helminth; D015801:Helminth Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D006801:Humans; D007124:Immunoenzyme Techniques; D008842:Microfilariae / Q000737:chemistry; D009854:Onchocerca / Q000737:chemistry; D017181:Onchocerca volvulus / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D013045:Species Specificity
D015801:Helminth Proteins; C114959:PXN-2 protein, Onchocerca volvulus; D011994:Recombinant Proteins