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Journal of Bacteriology, December 2008, p. 7904-7909, Vol. 190, No. 24
0021-9193/08/$08.00+0     doi:10.1128/JB.01116-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Direct Observation of Staphylococcus aureus Cell Wall Digestion by Lysostaphin {triangledown}

Grégory Francius,1 Oscar Domenech,2 Marie Paule Mingeot-Leclercq,2 and Yves F. Dufrêne1*

Unité de Chimie des Interfaces, Université Catholique de Louvain, Croix du Sud 2/18, B-1348 Louvain-la-Neuve, Belgium,1 Unité de Pharmacologie Cellulaire et Moléculaire, Université Catholique de Louvain, Avenue E. Mounier 73, B-1200 Brussels, Belgium2

Received 8 August 2008/ Accepted 25 September 2008

The advent of Staphylococcus aureus strains that are resistant to virtually all antibiotics has increased the need for new antistaphylococcal agents. An example of such a potential therapeutic is lysostaphin, an enzyme that specifically cleaves the S. aureus peptidoglycan, thereby lysing the bacteria. Here we tracked over time the structural and physical dynamics of single S. aureus cells exposed to lysostaphin, using atomic force microscopy. Topographic images of native cells revealed a smooth surface morphology decorated with concentric rings attributed to newly formed peptidoglycan. Time-lapse images collected following addition of lysostaphin revealed major structural changes in the form of cell swelling, splitting of the septum, and creation of nanoscale perforations. Notably, treatment of the cells with lysostaphin was also found to decrease the bacterial spring constant and the cell wall stiffness, demonstrating that structural changes were correlated with major differences in cell wall nanomechanical properties. We interpret these modifications as resulting from the digestion of peptidoglycan by lysostaphin, eventually leading to the formation of osmotically fragile cells. This study provides new insight into the lytic activity of lysostaphin and offers promising prospects for the study of new antistaphylococcal agents.

* Corresponding author. Mailing address: Unité de Chimie des Interfaces, Université Catholique de Louvain, Croix du Sud 2/18, B-1348 Louvain-la-Neuve, Belgium. Phone: (32) 10 47 36 00. Fax: (32) 10 47 20 05. E-mail: yves.dufrene{at}uclouvain.be

{triangledown} Published ahead of print on 3 October 2008.

Journal of Bacteriology, December 2008, p. 7904-7909, Vol. 190, No. 24
0021-9193/08/$08.00+0     doi:10.1128/JB.01116-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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