PARTIAL PURIFICATION AND CHARACTERIZATION OF LECTIN-LIKE ACTIVITIES FROM Lupinus albus SEEDS
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Instituto Nacional de Ciencias Agrícolas
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In our previous work, the haemagglutinating activity present on three Lupinus species was studied; from those results it was decided to purify lectin-like activities present in Lupinus albus seeds, taking into account the highest specificity previously found in the extracts. In this study, three appropriate affinity chromatography columns were prepared, in order to purify the haemagglutinating activity from L. albus. Pellet collected by centrifugation after saturating the extract with 80 % of ammonium sulphate was applied to Mucin- Sepharose, Fetuin-Sepharose and Galactose-Agarose columns respectively. Although lectin-like activity equivalent to about 5 % of the total protein in the extract was purified using the Mucin or Fetuin columns, the major haemagglutinating activity ran through and did not bind to any column tested. Proteins collected in the eluting peaks upon affinity chromatography were partially characterized by SDS electrophoresis and gel filtration in a Superose 12 column, in order to know about monomeric or oligomeric composition as well as their molecular weight. Circular dichroism spectra were also recorded to analyze the secondary structure of the protein.
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