ATP diphosphohydrolase from Schistosoma mansoni egg: characterization and immunocytochemical localization of a new antigen.
The fact that the Schistosoma mansoni egg has two ATP diphosphohydrolase (EC 3.6.1.5) isoforms with different net charges and an identical molecular weight of 63,000, identified by non-denaturing polyacrylamide gel electrophoresis and immunological cross-reactivity with potato apyrase antibodies, is shown. In soluble egg antigen (SEA), only the isoform with the lower net negative charge was detected and seemed to be the predominant species in this preparation. By confocal fluorescence microscopy, using anti-potato apyrase antibodies, the S. mansoni egg ATP diphosphohydrolase was detected on the external surface of miracidium and in von Lichtenberg's envelope. Intense fluorescence was also seen in the outer side of the egg-shell, entrapped by the surface microspines, suggesting that a soluble isoform is secreted. ATP diphosphohydrolase antigenicity was tested using the vegetable protein as antigen. The purified potato apyrase was recognized in Western blots by antibodies present in sera from experimentally S. mansoni-infected mice. In addition, high levels of IgG anti-ATP diphosphohydrolase antibodies were detected by ELISA in the same sera. This work represents the first demonstration of antigenic properties of S. mansoni ATP diphosphohydrolase and immunological cross-reactivity between potato apyrase and sera from infected individuals.
Faria-Pinto
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Departamento de Bioquímic/ICB, Universidade Federal de Juiz de Fora, 36015-400 Juiz de Fora, MG, Brasil.
Meirelles
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Lenzi
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Mota
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Penido
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Coelho
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Vasconcelos
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D000818:Animals; D000947:Antigens, Helminth / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D001081:Apyrase / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D007150:Immunohistochemistry; D007527:Isoenzymes; D008099:Liver / Q000469:parasitology; D051379:Mice; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D012550:Schistosoma mansoni / Q000201:enzymology* / Q000276:immunology / Q000378:metabolism
D000947:Antigens, Helminth; D007527:Isoenzymes; D001081:Apyrase