AFM Identifies a Protein Complex Involved in Pathogen Adhesion Which Ruptures at Three Nanonewtons

Chantraine, Constance; Mathelié‐Guinlet, Marion; Pietrocola, Giampiero; Speziale, Pietro; Dufrêne, Yves F.

doi:10.1021/acs.nanolett.1c02105

Cited by 10 publications

(7 citation statements)

References 48 publications

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“…4 and Appendix in Supplementary Material, Fig. S2 ) ( 32–34 ). Whereas rupture forces and binding probability were not altered for the WT–WT pairs (Fig.…”

Section: Resultsmentioning

confidence: 99%

The staphylococcal biofilm protein Aap mediates cell–cell adhesion through mechanically distinct homophilic and lectin interactions

et al. 2022

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The accumulation phase of staphylococcal biofilms relies both on the production of an extracellular polysaccharide matrix and on the expression of bacterial surface proteins. A prototypical example of such adhesive proteins is the long multidomain protein Aap from Staphylococcus epidermidis which mediates zinc-dependent homophilic interactions between Aap B-repeat regions through molecular forces that have not been investigated yet. Here, we unravel the remarkable mechanical strength of single Aap-Aap homophilic bonds between living bacteria and we demonstrate that intercellular adhesion also involves sugar binding through the lectin domain of the Aap A region. We find that the mechanical force needed to unfold individual β-sheet-rich G5-E domains from the Aap B-repeat regions is very high, ranging from 300 pN up to 1,000 pN at high loading rates, indicating these are extremely stable. This high mechanostability provides a means to the cells to form highly adhesive and cohesive biofilms capable to sustain high physiological shear stress. Importantly, we identify a previously undescribed role of Aap in bacterial-bacterial adhesion, that is, heterophilic sugar-binding by a specific lectin domain located in the N-terminal A-region, which might be important to establish initial contacts between cells before strong homophilic bonds come into play. This study emphasizes the remarkable mechanical and binding properties of Aap as well as its wide diversity of adhesive functions.

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“…4 and Appendix in Supplementary Material, Fig. S2 ) ( 32–34 ). Whereas rupture forces and binding probability were not altered for the WT–WT pairs (Fig.…”

Section: Resultsmentioning

confidence: 99%

The staphylococcal biofilm protein Aap mediates cell–cell adhesion through mechanically distinct homophilic and lectin interactions

et al. 2022

Self Cite

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“…AFM offers exceptional lateral resolution (up to nanometers), facilitating the visualization and characterization of individual molecules, [28][29][30] which can be employed to monitor molecular interactions under physiological conditions. [31][32][33][34][35][36] We previously developed a novel AFM system using a hybrid binding domain (HBD) to detect and quantify individual miRNAs within single cells.…”

Section: Resultsmentioning

confidence: 99%

AFM Imaging Reveals MicroRNA‐132 to be a Positive Regulator of Synaptic Functions

Park,

Kim,

Shin

et al. 2024

Advanced Science

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The modification of synaptic and neural connections in adults, including the formation and removal of synapses, depends on activity‐dependent synaptic and structural plasticity. MicroRNAs (miRNAs) play crucial roles in regulating these changes by targeting specific genes and regulating their expression. The fact that somatic and dendritic activity in neurons often occurs asynchronously highlights the need for spatial and dynamic regulation of protein synthesis in specific milieu and cellular loci. MicroRNAs, which can show distinct patterns of enrichment, help to establish the localized distribution of plasticity‐related proteins. The recent study using atomic force microscopy (AFM)‐based nanoscale imaging reveals that the abundance of miRNA(miR)‐134 is inversely correlated with the functional activity of dendritic spine structures. However, the miRNAs that are selectively upregulated in potentiated synapses, and which can thereby support prospective changes in synaptic efficacy, remain largely unknown. Using AFM force imaging, significant increases in miR‐132 in the dendritic regions abutting functionally‐active spines is discovered. This study provides evidence for miR‐132 as a novel positive miRNA regulator residing in dendritic shafts, and also suggests that activity‐dependent miRNAs localized in distinct sub‐compartments of neurons play bi‐directional roles in controlling synaptic transmission and synaptic plasticity.

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“…This knowledge can be applied to the treatment or prevention of bacterial infections [84]. AFM is also a powerful tool to study the interaction of MOs and host cells [85] [86]. By using singlemolecule AFM analysis [86] demostrates the affinity of S. aureus (adhesins) to endothelial cell integrins under different conditions (Low or high stress; 100 pN or 1000 -2000 pN).…”

Section: Microbiologymentioning

confidence: 99%

Holotomography and Atomic Force Microscopy: A Powerful Combination to Enhance Cancer, Microbiology and Nanotoxicology Research

Medina-Ramirez,

Macías-Díaz,

Masuoka

et al. 2023

Preprint

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Modern imaging strategies are paramount to studying living systems such as cells, bacteria, and fungi and their response to pathogens, toxicants, and nanomaterials as modulated by exposure and environmental factors. The need to understand the processes and mechanisms of damage, healing and cell survivability of living systems continues to motivate the development of alternative imaging strategies. Of particular interest is the use of label-free techniques that minimize interference of biological processes by foreign marking substances and reduce intense light exposure and potential photo-toxicity effects. This review focus on the potential synergic capabilities of atomic force microscopy (AFM) as a well-developed and robust imaging strategy with demonstrated applications to unravel intimate details in biomedical applications, with the label-free, fast, and enduring Holotomographic Microscopy (HT) strategy. We first review the operating principles that form the basis for the complementary details provided by these techniques regarding the surface and internal information provided by HT and AFM is essential and complimentary for the development of several biomedical areas studying the interaction mechanisms of nanomaterials with living organisms. First, AFM can provide superb resolution on surface morphology and biomechanical characterization. Second, the quantitative phase capabilities of HT enable superb modeling and quantification of the volume, surface area, protein content, and mass density of the main components of cells and microorganisms, including morphology of cells in microbiological systems. These capabilities result from directly quantifying refractive index changes without requiring fluorescent markers or chemicals. As such, HT is ideally suited for long-term monitoring of living organisms in conditions close to their natural settings. We present a case-based review of the principal uses of both techniques and their essential contributions to nanomedicine and nanotoxicology, emphasizing cancer and infectious disease control. The synergic impact of the sequential use of these complementary strategies provides a clear drive for adopting these techniques as interdependent fundamental tools.

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AFM Identifies a Protein Complex Involved in Pathogen Adhesion Which Ruptures at Three Nanonewtons

Cited by 10 publications

References 48 publications

The staphylococcal biofilm protein Aap mediates cell–cell adhesion through mechanically distinct homophilic and lectin interactions

The staphylococcal biofilm protein Aap mediates cell–cell adhesion through mechanically distinct homophilic and lectin interactions

AFM Imaging Reveals MicroRNA‐132 to be a Positive Regulator of Synaptic Functions

Holotomography and Atomic Force Microscopy: A Powerful Combination to Enhance Cancer, Microbiology and Nanotoxicology Research

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