Profiling the Bordetella pertussis Proteome during Iron Starvation

Abstract

Regulation of gene expression in response to local iron concentration is commonly observed in bacterial pathogens that face this nutrient limitation during host infection. In this study, a proteomic approach was used to analyze the differential protein expression of Bordetella pertussis under iron limitation. Whole cell lysates (WCL) and outer membrane fractions of bacteria grown either under iron-starvation or iron-excess conditions were analyzed by two-dimensional (2-D) gel electrophoresis. Statistical analysis revealed 36 proteins displaying differential expression, 9 with higher expression under iron-excess and 27 with increased expression under iron-starvation. These proteins were subjected to tryptic digestion and MALDI-TOF MS. Apart from those previously reported, we identified new low-iron-induced proteins that might help to explain the increased virulence of this phenotype. Additionally, we found evidence that at least one of the identified proteins, solely expressed under iron starvation, is highly immunogenic in infected individuals.

Characterization of co-purified acellular pertussis vaccines

“…23 Proteome of B. pertussis based on 2-DE have been studied to clarify the pathogenesis and immune mechanisms of pertussis. 24,25 In this study, proteomic technology was applied to clarify the composition of the co-purified APVs and compared the consistency of products between different batches and between different manufactures. A 2-DE protocol was developed to effectively resolve the protein mixture of the samples and subsequently protein spots were identified by MALDI-TOF/TOF MS.…”

Immunoproteomic Analysis of Human Serological Antibody Responses to Vaccination with Whole-Cell Pertussis Vaccine (WCV)

“…LivJ representing the most abundant transport protein has been characterized in the extracellular proteomes of E.coli BL21 and W3110 strains, showing that LivJ could also serve as extracellular protein released outside the cell and thus recognized by host immune system [79]. In addition to presence in the Surfaceome and OMV proteome of B. p , putative ABC transport solute binding protein as a newly identified low-iron-induced protein displays differential protein expression levels between iron-excess and iron-starvation conditions, and it might be closely associated with B.p virulence [80]. Furthermore, putative ABC transport solute binding protein of Neisseria meningitidis has been regarded as a potentially new protective antigen for incorporation into multicomponent meningococcal vaccines; it was not only found to be exposed on the bacterial surface and well conserved across a range of meningococcal circulating strains, but also strongly reactive with sera from 31 convalescent young children infected with meningococcal disease [23], [81].…”

High-Throughput Bioluminescence-Based Mutant Screening Strategy for Identification of Bacterial Virulence Genes

“…Identification of bacterial genes and proteins that are differentially expressed in response to a host defense can be an effective method for identifying putative bacterial virulence factors (4,9,48). In addition to microarrays and proteomics, other high-throughput methods have been used to identify bacterial genes upregulated in response to phagocytosis, including differential fluorescence induction (47), random luciferase transcriptional fusions (40), and selective capture of transcribed sequences (14,17).…”