CaracterizaÃÃo molecular da lectina ligadora de manose (MBL) em indivÃduos com hepatite C
AUTOR(ES)
Igor Teixeira Cavalcanti
DATA DE PUBLICAÇÃO
2007
RESUMO
Mannan-binding lectin (MBL), a member of the type-C lectins, is able to bind through multiple sites to various carbohydrate structures and activate complement. Low levels of MBL have been linked with susceptibility to several infectious diseases. Different assays used to MBL determination havenât been able to identify widely its molecular forms in the circulation. This work aimed to evaluate the molecular profile of MBL in serum of hepatitis C patients using lectin separation assay in microplates with nitrocellulose discs followed by immunorecognition and SDS-PAGE (reducing and non-reducing) 10% and Western blot analysis. The separation assay results showed a good binding efficiency of mannan-coated membrane and the MBL identification was confirmed to all genotypes by conventional dot-ELISA. Generally, few bands were visualized in samples submitted to separation assay, which might be correlated with very low MBL amount. The bands revealed some differences for the migration pattern between AA, AO and OO genotypes, showing detected marks in the range 50-90 kDa and 180 kDa, equivalents to low molecular mass variants of MBL structural subunits and high molecular mass, respectively. Non-reducing SDS-PAGE showed a greater range of molecular species, such as trimers (3x3), dimers and structural units MBL bands in some samples, mainly in AA individuals. The Western blot analysis confirmed the presence of high and low molecular mass similarly observed in the SDS-PAGE, with highlight the detection of 128 and 32 kDa MBL forms. This results suggests a simple and low coast separation method for MBL in which it was possible to evaluate the profile of serum forms of this lectin in Hepatitis C patients. This is an important step for the understanding the structure/genotype of MBL in this pathology which could be associated to the response to treatment
ASSUNTO(S)
electrophoresis lectina ligadora de manose eletroforese mbl bioquimica mannan-binding lectin mbl protein proteÃna
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