Characterization of the Burkholderia cenocepacia J2315 Surface-Exposed Immunoproteome
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Author
Sousa, Sílvia A.
Seixas, António M.M.
Mandal, Manoj
Rodríguez-Ortega, Manuel J.
Leitão, Jorge H.
Publisher
MDPIDate
2020Subject
Burkholderia cepacia complexSurface-exposed moieties
Liquid chromatography–tandem mass spectrometry (LC-MS/MS)
Immunoreactive proteins
Cystic fibrosis
Serum samples
Immunoreactivity
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Infections by the Burkholderia cepacia complex (Bcc) remain seriously life threatening to cystic fibrosis (CF) patients, and no effective eradication is available. A vaccine to protect patients against Bcc infections is a highly attractive therapeutic option, but none is available. A strategy combining the bioinformatics identification of putative surface-exposed proteins with an experimental approach encompassing the “shaving” of surface-exposed proteins with trypsin followed by peptide identification by liquid chromatography and mass spectrometry is here reported. The methodology allowed the bioinformatics identification of 263 potentially surface-exposed proteins, 16 of them also experimentally identified by the “shaving” approach. Of the proteins identified, 143 have a high probability of containing B-cell epitopes that are surface-exposed. The immunogenicity of three of these proteins was demonstrated using serum samples from Bcc-infected CF patients and Western blotting, validating the usefulness of this methodology in identifying potentially immunogenic surface-exposed proteins that might be used for the development of Bcc-protective vaccines.