American Chemical Society
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Tryptic Shaving of Staphylococcus aureus Unveils Immunodominant Epitopes on the Bacterial Cell Surface

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journal contribution
posted on 2020-06-25, 16:10 authored by Annette Dreisbach, Min Wang, Magdalena M. van der Kooi-Pol, Ewoud Reilman, Dennis G. A. M. Koedijk, Ruben A. T. Mars, José Duipmans, Marcel Jonkman, Joris J. Benschop, Hendrik P. J. Bonarius, Herman Groen, Michael Hecker, Andreas Otto, Katrin Bäsell, Jörg Bernhardt, Jaap Willem Back, Dörte Becher, Girbe Buist, Jan Maarten van Dijl
The opportunistic pathogen Staphylococcus aureus has become a major threat for human health and well-being by developing resistance to antibiotics and by fast evolution into new lineages that rapidly spread within the healthy human population. This calls for development of active or passive immunization strategies to prevent or treat acute phase infections. Since no such anti-staphylococcal immunization approaches are available for clinical implementation, the present studies were aimed at identifying new leads for their development. For this purpose, we profiled the cell-surface-exposed staphylococcal proteome under infection-mimicking conditions by combining two approaches for “bacterial shaving” with immobilized or soluble trypsin and subsequent mass spectrometry analysis of liberated peptides. In parallel, non-covalently cell-wall-bound proteins extracted with potassium thiocyanate and the exoproteome fraction were analyzed by gel-free proteomics. All data are available through ProteomeXchange accession PXD000156. To pinpoint immunodominant bacterial-surface-exposed epitopes, we screened selected cell-wall-attached proteins of S. aureus for binding of immunoglobulin G from patients who have been challenged by different types of S. aureus due to chronic wound colonization. The combined results of these analyses highlight particular cell-surface-exposed S. aureus proteins with highly immunogenic exposed epitopes as potential targets for development of protective anti-staphylococcal immunization strategies.