Accéder au contenu
Merck

Nanoscale adhesion forces of Pseudomonas aeruginosa type IV Pili.

ACS nano (2014-10-07)
Audrey Beaussart, Amy E Baker, Sherry L Kuchma, Sofiane El-Kirat-Chatel, George A O'Toole, Yves F Dufrêne
RÉSUMÉ

A variety of bacterial pathogens use nanoscale protein fibers called type IV pili to mediate cell adhesion, a primary step leading to infection. Currently, how these nanofibers respond to mechanical stimuli and how this response is used to control adhesion is poorly understood. Here, we use atomic force microscopy techniques to quantify the forces guiding the adhesion of Pseudomonas aeruginosa type IV pili to surfaces. Using chemical force microscopy and single-cell force spectroscopy, we show that pili strongly bind to hydrophobic surfaces in a time-dependent manner, while they weakly bind to hydrophilic surfaces. Individual nanofibers are capable of withstanding forces up to 250 pN, thereby explaining how they can resist mechanical stress. Pulling on individual pili yields constant force plateaus, presumably reflecting conformational changes, as well as nanospring properties that may help bacteria to withstand physiological shear forces. Analysis of mutant strains demonstrates that these mechanical responses originate solely from type IV pili, while flagella and the cell surface localized and proposed pili-associated adhesin PilY1 play no direct role. We also demonstrate that bacterial-host interactions involve constant force plateaus, the extension of bacterial pili, and the formation of membrane tethers from host cells. We postulate that the unique mechanical responses of type IV pili unravelled here enable the bacteria to firmly attach to biotic and abiotic surfaces and thus maintain attachment when subjected to high shear forces under physiological conditions, helping to explain why pili play a critical role in colonization of the host.

MATÉRIAUX
Référence du produit
Marque
Description du produit

Sigma-Aldrich
Phosphate de potassium dibasic, ACS reagent, ≥98%
Sigma-Aldrich
sulfate de magnésium, anhydrous, ReagentPlus®, ≥99.5%
Sigma-Aldrich
1-Dodecanethiol, ≥98%
Sigma-Aldrich
sulfate de magnésium, anhydrous, free-flowing, Redi-Dri, ReagentPlus®, ≥99.5%
Sigma-Aldrich
sulfate de magnésium, BioReagent, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
sulfate de magnésium, puriss. p.a., drying agent, anhydrous, ≥98.0% (KT), powder (very fine)
Sigma-Aldrich
Phosphate de potassium dibasic, anhydrous, for luminescence, for molecular biology, BioUltra, ≥99.0% (T)
Sigma-Aldrich
Magnesium sulfate solution, for molecular biology, 1.00 M±0.04 M
Sigma-Aldrich
sulfate de magnésium, anhydrous, free-flowing, Redi-Dri, reagent grade, ≥97%
Sigma-Aldrich
Potassium phosphate dibasic solution, 1.0 M
Sigma-Aldrich
Magnesium sulfate solution, BioUltra, for molecular biology
Sigma-Aldrich
sulfate de magnésium, ≥99.99% trace metals basis
Sigma-Aldrich
Phosphate de potassium dibasic, 99.95% trace metals basis
Sigma-Aldrich
Phosphate de potassium dibasic, reagent grade, ≥98.0%
Sigma-Aldrich
Potassium phosphate dibasic, meets USP testing specifications