Recombinant expression, purification and biochemical characterization of a superoxide dismutase from Entamoeba histolytica.
A recombinant iron-containing superoxide dismutase (recFeSOD) of Entamoeba histolytica was produced in a prokaryotic expression system. Purified recFeSOD was found to be enzymatically active as determined by (i) inhibition of ferri-cytochrome c reduction, (ii) dismutation of superoxide anions generated by human neutrophils and (iii) inhibition of nitroblue tetrazolium reduction. The enzymatic properties of recFeSOD were similar to those of the native protein in trophozoite extracts. In an ELISA using recFeSOD as antigen, 96% of sera from patients having invasive amebiasis were reactive whereas none of the healthy controls or of patients suffering from malaria, bacterial or viral infections were reactive. Only sera of Toxoplasma-, Leishmania- or Trypanosoma-infected individuals exhibited partial cross-reactivity to recFeSOD.
Bruchhaus
I
I
Bernhard Nocht Institute for Tropical Medicine, Hamburg, FR, Germany.
Brattig
N W
NW
Tannich
E
E
D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000953:Antigens, Protozoan / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D003429:Cross Reactions; D003574:Cytochrome c Group / Q000378:metabolism; D004748:Entamoeba histolytica / Q000201:enzymology* / Q000235:genetics / Q000276:immunology; D004749:Entamoebiasis / Q000097:blood / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007239:Infections / Q000097:blood / Q000276:immunology; D007891:Leishmania / Q000276:immunology; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000378:metabolism; D009580:Nitroblue Tetrazolium / Q000378:metabolism; D010084:Oxidation-Reduction; D015800:Protozoan Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D013482:Superoxide Dismutase / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D014122:Toxoplasma / Q000276:immunology; D014345:Trypanosoma / Q000276:immunology
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D003574:Cytochrome c Group; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D009580:Nitroblue Tetrazolium; D013482:Superoxide Dismutase