The crystal structure of Pneumolysin at 2.0 Å resolution reveals the molecular packing of the pre-pore complex

Pneumolysin is a cholesterol-dependent cytolysin (CDC) and virulence factor of Streptococcus pneumoniae. It kills cells by forming pores assembled from oligomeric rings in cholesterol-containing membranes. Cryo-EM has revealed the structures of the membrane-surface bound pre-pore and inserted-pore oligomers, however the molecular contacts that mediate these oligomers are unknown because high-resolution information is not available. Here we have determined the crystal structure of full-length pneumolysin at 1.98 Å resolution. In the structure, crystal contacts demonstrate the likely interactions that enable polymerisation on the cell membrane and the molecular packing of the pre-pore complex. The hemolytic activity is abrogated in mutants that disrupt these intermolecular contacts, highlighting their importance during pore formation. An additional crystal structure of the membrane-binding domain alone suggests that changes in the conformation of a tryptophan rich-loop at the base of the toxin promote monomer-monomer interactions upon membrane binding by creating new contacts. Notably, residues at the interface are conserved in other members of the CDC family, suggesting a common mechanism for pore and pre-pore assembly

Changes of pore structure and chloride content in cement pastes after pore solution expression

Pore solution expression is a widely accepted approach to extract pore solution of cement-based materials by appllying high pressure. In this study, the variations of pore solution distribution and chloride content in cement pastes before and after pore solution expression were examined. The results showed that the value of chloride concentration index N-c were mostly higher than 1.0 for cement pastes immersed in NaCl solution, and decreased with the chloride concentration of soaking solution and water-to-binder (w/b) ratio. During the pore solution expression, the pores larger than 40 nm were totally removed and the porosity of smaller pore was decreased. Based on a proposed physical model on structure of cement paste, the value of N-c was calculated according to the variations of pore structure and chloride content during pore solution expression. The calculated results showed similar trend as the experimental results obtained by pore solution expression method

Examining the Effect of Pore Size Distribution and Shape on Flow through Unsaturated Peat using Computer Tomography

The hydraulic conductivity of unsaturated peat soil is controlled by the air-filled porosity, pore size and geometric distribution as well as other physical properties of peat materials. This study investigates how the size and shape of pores affects the flow of water through peat soils. In this study we used X-ray Computed Tomography (CT), at 45μm resolution under 5 specific soil-water pressure head levels to provide 3-D, high-resolution images that were used to detect the inner pore structure of peat samples under a changing water regime. Pore structure and configuration were found to be irregular, which affected the rate of water transmission through peat soils. The 3-D analysis suggested that pore distribution is dominated by a single large pore-space. At low pressure head, this single large air-filled pore imparted a more effective flowpath compared to smaller pores. Smaller pores were disconnected and the flowpath was more tortuous than in the single large air-filled pore, and their contribution to flow was negligible when the single large pore was active. We quantify the pore structure of peat soil that affects the hydraulic conductivity in the unsaturated condition, and demonstrate the validity of our estimation of peat unsaturated hydraulic conductivity by making a comparison with a standard permeameter-based method. Estimates of unsaturated hydraulic conductivities were made for the purpose of testing the sensitivity of pore shape and geometry parameters on the hydraulic properties of peats and how to evaluate the structure of the peat and its affects on parameterization. We also studied the ability to quantify these factors for different soil moisture contents in order to define how the factors controlling the shape coefficient vary with changes in soil water pressure head. The relation between measured and estimated unsaturated hydraulic conductivity at various heads shows that rapid initial drainage, that changes the air-filled pore properties, creates a sharp decline in hydraulic conductivity. This is because the large pores readily lose water, the peat rapidly becomes less conductive and the flow path among pores, more tortuous

Layer Analysis of the Structure of Water Confined in Vycor Glass

A Molecular Dynamics simulation of the microscopic structure of water confined in a silica pore is presented. A single cavity in the silica glass has been modeled as to reproduce the main features of the pores of real Vycor glass. A layer analysis of the site-site radial distribution functions evidence the presence in the pore of two subsets of water molecules with different microscopic structure. Molecules which reside in the inner layer, close to the center of the pore, have the same structure as bulk water but at a temperature of 30 K higher. On the contrary the structure of the water molecules in the outer layer, close to the substrate, is strongly influenced by the water-substrate hydrophilic interaction and sensible distortions of the H-bond network and of the orientational correlations between neighboring molecules show up. Lowering the hydration has little effect on the structure of water in the outer layer. The consequences on experimental determinations of the structural properties of water in confinement are discussed.Comment: 6 pages, 8 figures included in the text, one figure added, changes in the tex

Reversible Self-Actuated Thermo-Responsive Pore Membrane.

Smart membranes, which can selectively control the transfer of light, air, humidity and temperature, are important to achieve indoor climate regulation. Even though reversible self-actuation of smart membranes is desirable in large-scale, reversible self-regulation remains challenging. Specifically, reversible 100% opening/closing of pore actuation showing accurate responsiveness, reproducibility and structural flexibility, including uniform structure assembly, is currently very difficult. Here, we report a reversible, thermo-responsive self-activated pore membrane that achieves opening and closing of pores. The reversible, self-actuated thermo-responsive pore membrane was fabricated with hybrid materials of poly (N-isopropylacrylamide), (PNIPAM) within polytetrafluoroethylene (PTFE) to form a multi-dimensional pore array. Using Multiphysics simulation of heat transfer and structural mechanics based on finite element analysis, we demonstrated that pore opening and closing dynamics can be self-activated at environmentally relevant temperatures. Temperature cycle characterizations of the pore structure revealed 100% opening ratio at T = 40 °C and 0% opening ratio at T = 20 °C. The flexibility of the membrane showed an accurate temperature-responsive function at a maximum bending angle of 45°. Addressing the importance of self-regulation, this reversible self-actuated thermo-responsive pore membrane will advance the development of future large-scale smart membranes needed for sustainable indoor climate control

Structural insights into Clostridium perfringens delta toxin pore formation

Clostridium perfringens Delta toxin is one of the three hemolysin-like proteins produced by C. perfringens type C and possibly type B strains. One of the others, NetB, has been shown to be the major cause of Avian Nectrotic Enteritis, which following the reduction in use of antibiotics as growth promoters, has become an emerging disease of industrial poultry. Delta toxin itself is cytotoxic to the wide range of human and animal macrophages and platelets that present GM2 ganglioside on their membranes. It has sequence similarity with Staphylococcus aureus β-pore forming toxins and is expected to heptamerize and form pores in the lipid bilayer of host cell membranes. Nevertheless, its exact mode of action remains undetermined. Here we report the 2.4 Å crystal structure of monomeric Delta toxin. The superposition of this structure with the structure of the phospholipid-bound F component of S. aureus leucocidin (LukF) revealed that the glycerol molecules bound to Delta toxin and the phospholipids in LukF are accommodated in the same hydrophobic clefts, corresponding to where the toxin is expected to latch onto the membrane, though the binding sites show significant differences. From structure-based sequence alignment with the known structure of staphylococcal α-hemolysin, a model of the Delta toxin pore form has been built. Using electron microscopy, we have validated our model and characterized the Delta toxin pore on liposomes. These results highlight both similarities and differences in the mechanism of Delta toxin (and by extension NetB) cytotoxicity from that of the staphylococcal pore-forming toxins