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Title: Identificação de metalotioneínas em ostra crassostrea rhizophorae (Guilding, 1828) para aplicações ambientais
Keywords: Biomonitoramento;  Biomarcadores;  Metalotioneínas;  Metaistraço;  Crassostrea rhizophorae;  Biomonitoring;  Biomarkers;  Metallothioneins;  Trace metals;  Crassostrea rhizophorae
Issue Date: 28-Jun-2007
Abstract: In this work metallothioneins (MTs) were identified in the mangrove oyster Crassostrea rhizophorae to be used in environmental studies, employing modifications in the protein extraction method described by Viarengo et al. (1997). The oyster s proteins were submitted to SDS-PAGE 16,5% and the MT s cysteine were marked by means of monobromobimane (mBBr) derivatization, immunoblotting with a specific MT oyster antibody and ESI ion trap mass spectrometry. In addition, an oyster group was exposed to Cd+2 in order to detect what protein bands would be induced by this metal. We also determined the concentrations of MT, Zn, Cd and Cu in native oysters from four different sites along the Rio de Janeiro coast. The adaptations in the original protocol improved the analyses, resulting in reproducible data. The mBBr and the antibody techniques identified mainly the same bands, as well as the MT standards. However, mass spectrometry provided no reliable results due to keratin contamination in the gels. Only the positive control (Bovine Serum Albumine) and the MT standard were recognized. Oysters exposed to cadmium exhibited induction of the 14 kDa protein band. The determination of MT 14 kDa, Zn, Cd and Cu in digestive glands of native oysters did not show significant correlations between this protein and metals. The lack of correlations between Zn, Cd and Cu and metallothioneins can be attributed to the kind of fraction analyzed in tissue. The adaptations in the protein extraction protocol, coupled with the mBBr derivatization, are promising techniques for routine analyses of environmental samples in biomonitoring studies employing MTs as biomarkers for metal contamination.
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