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Nanoscale mapping and functional analysis of individual adhesins on living bacteria

Nature Methods volume 2, pages 515–520 (2005)Cite this article

A Corrigendum to this article was published on 01 August 2005

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Abstract

Although much progress has been made in the identification and characterization of adhesins borne by pathogenic bacteria, the molecular details underlying their interaction with host receptors remain largely unknown owing to the lack of appropriate probing techniques. Here we report a method, based on atomic force microscopy (AFM) with tips bearing biologically active molecules, for measuring the specific binding forces of individual adhesins and for mapping their distribution on the surface of living bacteria. First, we determined the adhesion forces between the heparin-binding haemagglutinin adhesin (HBHA) produced by Mycobacterium tuberculosis and heparin, used as a model sulphated glycoconjugate receptor. Both the adhesion frequency and adhesion force increased with contact time, indicating that the HBHA-heparin complex is formed via multiple intermolecular bridges. We then mapped the distribution of single HBHA molecules on the surface of living mycobacteria and found that the adhesin is not randomly distributed over the mycobacterial surface, but concentrated into nanodomains.

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Figure 1: Strategy for measuring the HBHA-heparin binding force at the single-molecule level using AFM.
Figure 2: Measurement of the binding force of the HBHA-heparin complex.
Figure 3: Dynamics of the HBHA-heparin interaction.
Figure 4: Mapping single HBHA adhesins on living mycobacteria.
Figure 5: Distribution of HBHA on the surface of M. bovis BCG and M. bovis BCG ΔHBHA as probed using different tips and cell preparations.

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  • 22 June 2005

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Notes

  1. NOTE:The website listed in the article has been modified. The corrected citation should read: (F.D.M. and M.C. Vidal-Pessolani, unpublished data; http://genolist.pasteur.fr/Leproma/) The error has been corrected in the HTML version of the article, and this correction has been appended to the PDF version of the article.

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Acknowledgements

This work was supported by the National Foundation for Scientific Research (FNRS), the Université Catholique de Louvain (Fonds Spéciaux de Recherche), the Federal Office for Scientific, Technical and Cultural Affairs (Interuniversity Poles of Attraction Programme), the Research Department of the Communauté Française de Belgique (Concerted Research Action), the INSERM, the Institut Pasteur de Lille and the Région Nord-Pas de Calais. N.L.A. and B.H.C. acknowledge support from the US National Science Foundation through the MRSEC program. We thank G. Delogù for the gift of E. coli BL21(pGD51), E. Pradel and M. Simonet for critical reading of the manuscript, L. Piraux for the use of the thermal evaporator, E. Ferain for the use of the scanning electron microscope and P. Hinterdorfer for stimulating discussions. Y.F.D. is a Research Associate of the FNRS.

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Author notes

  1. Vincent Dupres and Franco D Menozzi: These authors contributed equally to this work.

Authors and Affiliations

  1. Unité de Chimie des Interfaces, Université Catholique de Louvain, Croix du Sud 2/18, Louvain-la-Neuve, B-1348, Belgium

    Vincent Dupres & Yves F Dufrêne

  2. INSERM Avenir, Pathogénie des Légionelles, Institut Pasteur de Lille, IFR17, 1 rue du Professeur Calmette, Lille, F-59019, Cedex, France

    Franco D Menozzi & Dominique Raze

  3. INSERM U629, Lille, Mécanismes Moléculaires de la Pathogénie Microbienne, Institut Pasteur de Lille, IFR17, 1 rue du Professeur Calmette, Lille, F-59019, Cedex, France

    Franco D Menozzi, Camille Locht & Dominique Raze

  4. Department of Chemical and Biological Engineering, University of Wisconsin-Madison, 1415 Engineering Drive, Madison, 53706, Wisconsin, USA

    Brian H Clare & Nicholas L Abbott

  5. Institut des Matériaux Jean Rouxel, Laboratoire de Physique des Matériaux et des Nanostructures, 2 rue de la Houssinière, Nantes, F-44322, Cedex 3, France

    Stéphane Cuenot

  6. CNRS UMR 8525, Institut de Biologie de Lille, 1 rue du Professeur Calmette, Lille, F-59019, Cedex, France

    Coralie Bompard

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  1. Vincent Dupres
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Correspondence to Franco D Menozzi or Yves F Dufrêne.

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Dupres, V., Menozzi, F., Locht, C. et al. Nanoscale mapping and functional analysis of individual adhesins on living bacteria. Nat Methods 2, 515–520 (2005). https://doi.org/10.1038/nmeth769

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