Lysostaphin is a bacteriolytic enzyme targeting peptidoglycan, the essential component of the bacterial cell envelope. It displays a very potent and specific activity toward staphylococci, including methicillin-resistant Staphylococcus aureus. Lysostaphin causes rapid cell lysis and disrupts biofilms, and is therefore a therapeutic agent of choice to eradicate staphylococcal infections. The C-terminal SH3b domain of lysostaphin recognizes peptidoglycans containing a pentaglycine crossbridge and has been proposed to drive the preferential digestion of staphylococcal cell walls. Here we elucidate the molecular mechanism underpinning recognition of staphylococcal peptidoglycan by the lysostaphin SH3b domain. We show that the pentaglycine crossbridge and the peptide stem are recognized by two independent binding sites located on opposite sides of the SH3b domain, thereby inducing a clustering of SH3b domains. We propose that this unusual binding mechanism allows synergistic and structurally dynamic recognition of S. aureus peptidoglycan and underpins the potent bacteriolytic activity of this enzyme.
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Structural data have been deposited in the PDB with coordinate accession numbers 6RK4 (high-resolution set) and 6RJE (home source set). All other data generated or analyzed during this study are included in this article and its supplementary information or are available from the corresponding authors upon request.
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L.S.G.-D. is a PhD student funded by the Mexican government through a CONACYT scholarship. H.W.-M. is supported by a BBSRC MIBTP studentship. We thank the BBSRC and EPSRC for funding to upgrade the 600- and 800-MHz spectrometers, respectively (grant numbers BB/R000727/1 and EP/S01358X/1). The work in the laboratory of I.S. is supported by the Foundation for Polish Science (FNP) program, co-financed by the European Union under the European Regional Development Fund (grant TEAMTECH/2016-3/19).
The authors declare no competing interests.
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Gonzalez-Delgado, L.S., Walters-Morgan, H., Salamaga, B. et al. Two-site recognition of Staphylococcus aureus peptidoglycan by lysostaphin SH3b. Nat Chem Biol 16, 24–30 (2020). https://doi.org/10.1038/s41589-019-0393-4
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