32 research outputs found

    A probabilistic model for crystal growth applied to protein deposition at the microscale

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    A probabilistic discrete model for 2D protein crystal growth is presented. This model takes into account the available space and can describe growing processes of different nature due to the versatility of its parameters which gives the model great flexibility. The accuracy of the simulation is tested against a real protein (SbpA) crystallization experiment showing high agreement between the proposed model and the actual images of the nucleation process. Finally, it is also discussed how the regularity of the interface (i.e. the curve that separates the crystal from the substrate) affects to the evolution of the simulation.Comment: 13 pages, 12 figure

    Analysis of the cell surface layer ultrastructure of the oral pathogen Tannerella forsythia

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    The Gram-negative oral pathogen Tannerella forsythia is decorated with a 2D crystalline surface (S-) layer, with two different S-layer glycoprotein species being present. Prompted by the predicted virulence potential of the S-layer, this study focused on the analysis of the arrangement of the individual S-layer glycoproteins by a combination of microscopic, genetic, and biochemical analyses. The two S-layer genes are transcribed into mRNA and expressed into protein in equal amounts. The S-layer was investigated on intact bacterial cells by transmission electron microscopy, by immune fluorescence microscopy, and by atomic force microscopy. The analyses of wild-type cells revealed a distinct square S-layer lattice with an overall lattice constant of 10.1 ± 0.7 nm. In contrast, a blurred lattice with a lattice constant of 9.0 nm was found on S-layer single-mutant cells. This together with in vitro self-assembly studies using purified (glyco)protein species indicated their increased structural flexibility after self-assembly and/or impaired self-assembly capability. In conjunction with TEM analyses of thin-sectioned cells, this study demonstrates the unusual case that two S-layer glycoproteins are co-assembled into a single S-layer. Additionally, flagella and pilus-like structures were observed on T. forsythia cells, which might impact the pathogenicity of this bacterium

    Fluorescence spectroscopy on polyelectrolyte free standing films

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    Influence of ethanol on the thickness and free energy of film formation of DMPC foam films

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    Foam films prepared from 1,2-dimirystoil-sn-glycero-3-phosphorylcholine (DMPC) dispersions in water–ethanol mixtures were investigated. Their thickness and contact angle (foam film/meniscus) were measured. Experimental results show that an increase of EtOH concentration in the film forming dispersions leads to a decrease in the film thickness. At EtOH concentrations above 40% v/v the foam films have a bilayer structure without any noticeable core of solvent. The film thickness remains constant with the further increasing of the EtOH concentration up until 50% v/v. This behaviour is corroborated by the strong increase in the contact angles (a decrease in the free energy of film formation) with increasing EtOH concentration. Ellipsometric measurements on the thickness of adsorbed DMPC monolayers and surface pressure isotherms of DMPC spread on the water–ethanol subphases show that the effective area per lipid molecule decreases, resulting in a larger monolayer thickness, when the EtOH concentration in the subphase is increased. An increase of the EtOH concentration leads to a dehydration of the DMPC molecules and a reduction in strength and range of the repulsive hydration force between the film monolayers. The film thickness and the free energy of film formation are governed by the balance of the van der Waals attraction and the repulsive hydration force between the foam film surfaces
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