Mapeamento e síntese de epitopos da mutalisina II (LHF-II) metaloproteinase purificada do veneno de Lachesis muta muta

Abstract

SPOT - synthesis technique has been used to construct immobilized linear peptides (twelve amino acids residues overlapping nine residues) over cellulose surface covering the entire primary amino acid sequence of mutalisin II, a metalloproteinase responsible for hemorrhagic activity of Lachesis muta muta venom. Rabbit serum against mutalisin II reacted with these peptides and revealed three major immunogenic regions: a C-terminal, a Central and N-terminal of the protein. A peptide sequence of each identified immunogenic region was selected for its chemical synthesis. The peptides were purified and analyzed by mass spectroscopy and these peptides were recognized by rabbit serumagainst mutalisin II in an ELISA assay. The peptides were coupled to KLH (Keyhole Limpet Hemocyanin) for the production of antipeptides antibodies in rabbit. These antibodies had a mild recognition of mutalisin II and a weak recognition of the whole venom. The IgG fraction of the rabbit serum antipeptides was purified for an inhibition assay of this enzyme. These antibodies were capable to inhibit the proteolitic activity ofthe enzyme by 63,5%.