Open AccessEpitope mapping of Burkholderia pseudomallei serine metalloprotease: identification of serine protease epitope mimics
Author(s) -
Chan ShzuWei,
Nathan Sheila
Publication year - 2005
Publication title -
fems immunology & medical microbiology
Language(s) - English
Resource type - Journals
eISSN - 1574-695X
pISSN - 0928-8244
DOI - 10.1016/j.femsim.2004.06.025
Subject(s) - polyclonal antibodies , epitope , burkholderia pseudomallei , linear epitope , biopanning , biology , serine protease , microbiology and biotechnology , protease , phage display , serine , metalloproteinase , peptide sequence , peptide , epitope mapping , peptide library , biochemistry , antibody , enzyme , bacteria , gene , genetics
Filamentous phage random peptide libraries were used to identify the epitopes of Burkholderia pseudomallei protease by panning against IgG polyclonal sera that exhibited protease neutralizing properties. The isolated fusion peptides presented a consensus peptide sequence, T K SMA L SG , which closely resembles part of the active site sequence, 435G TSMAT PHVA G 445, of B. pseudomallei serine metalloprotease. By comparing the consensus sequence, TKSMALSG, with the predicted three‐dimensional molecular model of B. pseudomallei serine metalloprotease, it appears that the potential antibody binding epitope was buried within the molecule. This active site was conformational whereby one continuous sub‐region (SMA) was located between two discontinuous sub‐regions, supplied by the flanking residues in the same polypeptide. All phages selected from the biopanning with IgG polyclonal sera showed good binding towards the polyclonal antibodies when compared to the negative control. In addition, these peptide‐bearing phages showed competitive inhibition of B. pseudomallei serine metalloprotease binding to the polyclonal IgG.