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Related Experiment Video

Updated: Jun 22, 2026

Investigating Receptor-ligand Systems of the Cellulosome with AFM-based Single-molecule Force Spectroscopy
11:34

Investigating Receptor-ligand Systems of the Cellulosome with AFM-based Single-molecule Force Spectroscopy

Published on: December 20, 2013

Stretching polysaccharides on live cells using single molecule force spectroscopy.

Grégory Francius1, David Alsteens, Vincent Dupres

  • 1Unité de Chimie des Interfaces, Université Catholique de Louvain, Louvain-la-Neuve, Belgium.

Nature Protocols
|May 30, 2009
PubMed
Summary
This summary is machine-generated.

This study introduces a single-molecule force spectroscopy (SMFS) protocol to analyze polysaccharide chains on live bacteria. The method allows detailed mapping of polysaccharide localization, adhesion, and mechanical properties.

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Area of Science:

  • Biophysics
  • Microbiology
  • Materials Science

Background:

  • Understanding cell surface molecule mechanics is crucial for function.
  • Single-molecule force spectroscopy (SMFS) enables probing of live cell adhesion and mechanics.
  • Bacterial surface polysaccharides play vital roles in cell interactions and environment adaptation.

Purpose of the Study:

  • To present a detailed protocol for analyzing polysaccharide chains on live bacteria using SMFS.
  • To demonstrate the application of SMFS for probing the mechanical and adhesive properties of bacterial polysaccharides.
  • To provide a framework for investigating the elasticity of individual polysaccharide chains.

Main Methods:

  • Functionalization of atomic force microscopy (AFM) tips with specific lectins (e.g., Pseudomonas aeruginosa-I, concanavalin A).
  • Utilizing SMFS to stretch individual polysaccharide chains on live Lactobacillus rhamnosus GG.
  • Mapping the localization, adhesion forces, and extension of polysaccharide molecules.
  • Applying the extended freely jointed chain model for data analysis of macromolecular elasticity.

Main Results:

  • The protocol successfully analyzes polysaccharide chains of different types on live bacteria.
  • SMFS with lectin-functionalized tips allows for the characterization of individual polysaccharide chain mechanics.
  • Data analysis provides insights into the elasticity of stretched polysaccharide macromolecules.
  • The protocol is adaptable for analyzing other cell types.

Conclusions:

  • The developed SMFS protocol offers a powerful tool for dissecting the mechanical and adhesive properties of bacterial cell surface polysaccharides.
  • This method advances the understanding of how polysaccharide structure influences bacterial adhesion and function.
  • The protocol's adaptability makes it valuable for broader applications in cell surface biomechanics research.