AFM Nanoindentation Experiments on Protein Shells: A Protocol

Guo, Yukun; Roos, Wouter H.

doi:10.1007/978-1-4939-8894-5_14

Cited by 8 publications

(9 citation statements)

References 46 publications

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“…To immobilize noroVLPs during AFM measurements, a hydrophobic glass cover slip was applied as a substrate. The hydrophobic coating on the glass cover slip was obtained by applying a hexamethyldisilazane (Sigma-Aldrich) treatment as described previously [ 29 , 30 ]. A droplet of 200 μL diluted noroVLP solution was deposited on a hydrophobic glass cover slip.…”

Section: Methodsmentioning

confidence: 99%

Fucose Binding Cancels out Mechanical Differences between Distinct Human Noroviruses

Feng

Pogan

Thiede

et al. 2023

Viruses

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The majority of nonbacterial gastroenteritis in humans and livestock is caused by noroviruses. Like most RNA viruses, frequent mutations result in various norovirus variants. The strain-dependent binding profiles of noroviruses to fucose are supposed to facilitate norovirus infection. It remains unclear, however, what the molecular mechanism behind strain-dependent functioning is. In this study, by applying atomic force microscopy (AFM) nanoindentation technology, we studied norovirus-like particles (noroVLPs) of three distinct human norovirus variants. We found differences in viral mechanical properties even between the norovirus variants from the same genogroup. The noroVLPs were then subjected to fucose treatment. Surprisingly, after fucose treatment, the previously found considerable differences in viral mechanical properties among these variants were diminished. We attribute a dynamic switch of the norovirus P domain upon fucose binding to the reduced differences in viral mechanical properties across the tested norovirus variants. These findings shed light on the mechanisms used by norovirus capsids to adapt to environmental changes and, possibly, increase cell infection. Hereby, a new step towards connecting viral mechanical properties to viral prevalence is taken.

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Section: Methodsmentioning

confidence: 99%

Fucose Binding Cancels out Mechanical Differences between Distinct Human Noroviruses

Feng

Pogan

Thiede

et al. 2023

Viruses

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“…Curves obtained by a sharp probe often had failures along the approaching part (Figure 4c). When studying viruses, protein shells and lipid bilayers, such loading curves usually were attributed to a rupture of a particle surface [27,28,29]. The sharp CSG10 tip resembles a needle in its geometry.…”

Section: Resultsmentioning

confidence: 99%

Assessment of Primary Cell Wall Nanomechanical Properties in Internal Cells of Non-Fixed Maize Roots

Kozlova

Petrova

Ананченко

et al. 2019

Plants

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The mechanical properties of cell walls play a vital role in plant development. Atomic-force microscopy (AFM) is widely used for characterization of these properties. However, only surface or isolated plant cells have been used for such investigations, at least as non-embedded samples. Theories that claim a restrictive role of a particular tissue in plant growth cannot be confirmed without direct measurement of the mechanical properties of internal tissue cell walls. Here we report an approach of assessing the nanomechanical properties of primary cell walls in the inner tissues of growing plant organs. The procedure does not include fixation, resin-embedding or drying of plant material. Vibratome-derived longitudinal and transverse sections of maize root were investigated by AFM in a liquid cell to track the changes of cell wall stiffness and elasticity accompanying elongation growth. Apparent Young’s modulus values and stiffness of stele periclinal cell walls in the elongation zone of maize root were lower than in the meristem, i.e., cell walls became more elastic and less resistant to an applied force during their elongation. The trend was confirmed using either a sharp or spherical probe. The availability of such a method may promote our understanding of individual tissue roles in the plant growth processes.

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“…Prior to AFM measurement, different HPV16 particles stock solutions were first diluted to a concentration of 11.3 μg/ml. Generally, AFM imaging and nanoindentation experiments were conducted following the previous protocol 65 . Briefly, a droplet of 100 μL HPV solution was deposited on a hydrophobic glass coverslip and incubated for 15 min at room temperature before adding another 1 ml PBS to a liquid receptacle.…”

Section: Methodsmentioning

confidence: 99%

Glycan-induced structural activation softens the human papillomavirus capsid for entry through reduction of intercapsomere flexibility

Feng,

van Bodegraven,

Kádek

et al. 2024

Preprint

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High-risk human papillomaviruses (HPVs) cause various cancers. While type-specific prophylactic vaccines are available, additional anti-viral strategies are highly desirable. Initial HPV cell entry involves receptor-switching induced by structural capsid modifications. These modifications are initiated by interactions with cellular heparan sulphates (HS), however, their molecular nature and functional consequences remain elusive. Combining virological assays with hydrogen/deuterium exchange mass spectrometry, and atomic force microscopy, we investigated the effect of capsid-HS binding and structural activation. We show how HS-induced structural activation requires a minimal HS-chain length and simultaneous engagement of several binding sites by a single HS molecule. This engagement introduces a pincer-like force that stabilizes the capsid in a conformation with extended capsomer linkers. It results in capsid enlargement and softening, thereby facilitating L1 proteolytic cleavage and subsequent L2-externalization, as needed for cell entry. Our data will help further devising prophylactic strategies against HPV infections.

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AFM Nanoindentation Experiments on Protein Shells: A Protocol

Cited by 8 publications

References 46 publications

Fucose Binding Cancels out Mechanical Differences between Distinct Human Noroviruses

Fucose Binding Cancels out Mechanical Differences between Distinct Human Noroviruses

Assessment of Primary Cell Wall Nanomechanical Properties in Internal Cells of Non-Fixed Maize Roots

Glycan-induced structural activation softens the human papillomavirus capsid for entry through reduction of intercapsomere flexibility

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