Title: Deciphering the Nanometer-Scale Organization and Assembly of Lactobacillus rhamnosus GG Pili using Atomic Force Microscopy
Authors: Tripathi, Prachi ×
Dupres, V.
Beaussart, A.
Lebeer, Sarah
Claes, Ingmar
Vanderleyden, Jos
Dufrêne, Y. #
Issue Date: 2012
Publisher: American Chemical Society
Series Title: Langmuir vol:28 pages:2211-2216
Abstract: In living cells, sophisticated functional interfaces are generated through the selfassembly
of bioactive building blocks. Prominent examples of such biofunctional surfaces are
bacterial nanostructures referred to as pili. Although these proteinaceous filaments exhibit
remarkable structure and functions, their potential to design bio-inspired self-assembled
systems has been overlooked. Here, we used atomic force microscopy (AFM) to explore the
supramolecular organization and self-assembly of pili from the Gram-positive probiotic
bacterium Lactobacillus rhamnosus GG (LGG). High-resolution AFM imaging of cell
preparations adsorbed on mica revealed pili not only all around the cells, but also in the
form of remarkable star-like structures assembled on the mica surface. Next, we showed
that two-step centrifugation is a simple procedure to separate large amounts of pili, even
though through their synthesis they are covalently anchored to the cell wall. We also found
that the centrifuged pili assemble as long bundles. We suggest that these bundles originate
from a complex interplay of mechanical effects (centrifugal force) and biomolecular
interactions involving the SpaC cell adhesion pilin subunit (lectin-glycan bonds, hydrophobic
bonds). Supporting this view, we found that pili isolated from an LGG mutant lacking
hydrophilic exopolysaccharides show an increased tendency to form tight bundles. These
experiments demonstrate that AFM is a powerful platform for visualizing individual pili on
bacterial surfaces and for unravelling their two-dimensional assembly on solid surfaces. Our
data suggest that bacterial pili may provide a generic approach in nanobiotechnology for
elaborating functional supramolecular interfaces assembled from bioactive building blocks.
ISSN: 0743-7463
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Centre of Microbial and Plant Genetics
× corresponding author
# (joint) last author

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