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Article
Nature Methods  2, 515 - 520 (2005)
Published online: 22 June 2005; Corrected online: 22 June 2005 | doi:10.1038/nmeth769


There is a Corrigendum (August 2005) associated with this Article.

Nanoscale mapping and functional analysis of individual adhesins on living bacteria

Vincent Dupres1, 7, Franco D Menozzi2, 3, 7, Camille Locht3, Brian H Clare4, Nicholas L Abbott4, Stéphane Cuenot5, Coralie Bompard6, Dominique Raze2, 3 & Yves F Dufrêne1

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

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

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

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

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

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

7  These authors contributed equally to this work.

Correspondence should be addressed to Yves F Dufrêne dufrene@cifa.ucl.ac.be or Franco D Menozzi franco.menozzi@ibl.fr
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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Nature Methods
ISSN: 1548-7091
EISSN: 1548-7105
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