In this study, we used gel-free nanoLC-MS/MS-based proteomics to compare the protein profile of B. pertussis wild type and an isogenic hfq defective mutant strain under control an iron limited conditions. We found that Hfq affects the abundance of 302 proteins, which represents 8% of the total B. pertussis coding sequence. The absence of Hfq induced changes in the abundance of proteins involved in metabolic pathways, stress response and virulence. Hfq was also found involved in B. pertussis adaptation to iron starvation, one of the main stresses this pathogen faces inside the host. Altogether, these results indicate that B. pertussis Hfq is involved in bacterial physiological processes as well as bacterial pathogenesis.