Abstract
New proteomic techniques have led to more sensitive approaches to ascertain differences between bacterial strains. Here we apply these methods and use a comparative proteomic analysis to determine if Bordetella pertussis (Bp) strains C056, D956, and F646 (C, D, F) show differences in the expression of membrane proteins compared to the laboratory strain Tohama I (T). Bp isolates were fractionated to enrich for membrane proteins, and subsequently analyzed using an electrospray tandem mass spectrometer. The data was mined using the Mascot search algorithm, and the results were processed to generate a validated list of 206 Bp proteins in the membrane fraction. Several proteins (including those found in the vaccine) were common to all strains. The strain comparisons also showed 31, 17, 18, and 11 proteins unique to T, C, D, and F respectively, including outer membrane lipoproteins, pertussis toxin liberation protein F, and serotype 2 fimbrial subunit precursor proteins. Immunoblots of the membrane fraction using murine antisera against all four strains or human antisera from culture confirmed patients revealed at least 1 unique immunogenic protein per strain. These proteins are under further investigation as potential candidates for diagnostic assay development and to determine if proteomic analysis can contribute to differences and similarities observed in epidemiologic investigations. Supported by APHL/CDC.
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