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Applied and Environmental Microbiology, December 2008, p. 7522-7528, Vol. 74, No. 24
0099-2240/08/$08.00+0     doi:10.1128/AEM.00948-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Staphylococcus aureus-Fibronectin Interactions with and without Fibronectin-Binding Proteins and Their Role in Adhesion and Desorption

Chun-Ping Xu,¹ Niels P. Boks,¹ Joop de Vries,¹ Hans J. Kaper,¹ Willem Norde,^1,2 Henk J. Busscher,¹ and Henny C. van der Mei^1*

Department of Biomedical Engineering, University Medical Center Groningen, and University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands,¹ Laboratory of Physical Chemistry and Colloid Science, Wageningen University, P.O. Box 8038, 6700 EK Wageningen, The Netherlands²

Received 25 April 2008/ Accepted 13 October 2008

Adhesion and residence-time-dependent desorption of two Staphylococcus aureus strains with and without fibronectin (Fn) binding proteins (FnBPs) on Fn-coated glass were compared under flow conditions. To obtain a better understanding of the role of Fn-FnBP binding, the adsorption enthalpies of Fn with staphylococcal cell surfaces were determined using isothermal titration calorimetry (ITC). Interaction forces between staphylococci and Fn coatings were measured using atomic force microscopy (AFM). The strain with FnBPs adhered faster and initially stronger to an Fn coating than the strain without FnBPs, and its Fn adsorption enthalpies were higher. The initial desorption was high for both strains but decreased substantially within 2 s. These time scales of staphylococcal bond ageing were confirmed by AFM adhesion force measurement. After exposure of either Fn coating or staphylococcal cell surfaces to bovine serum albumin (BSA), the adhesion of both strains to Fn coatings was reduced, suggesting that BSA suppresses not only nonspecific but also specific Fn-FnBP interactions. Adhesion forces and adsorption enthalpies were only slightly affected by BSA adsorption. This implies that under the mild contact conditions of convective diffusion in a flow chamber, adsorbed BSA prevents specific interactions but does allow forced Fn-FnBP binding during AFM or stirring in ITC. The bond strength energies calculated from retraction force-distance curves from AFM were orders of magnitude higher than those calculated from desorption data, confirming that a penetrating Fn-coated AFM tip probes multiple adhesins in the outermost cell surface that remain hidden during mild landing of an organism on an Fn-coated substratum, like that during convective diffusional flow.

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* Corresponding author. Mailing address: Department of Biomedical Engineering, University Medical Center Groningen, and University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands. Phone: 31-50-363-3140. Fax: 31-50-363-3159. E-mail: h.c.van.der.mei{at}med.umcg.nl

Published ahead of print on 24 October 2008.

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Applied and Environmental Microbiology, December 2008, p. 7522-7528, Vol. 74, No. 24
0099-2240/08/$08.00+0     doi:10.1128/AEM.00948-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.