High-resolution atomic force microscopy visualization of metalloproteins and their complexes

Background

Metalloproteins myeloperoxidase (MPO), ceruloplasmin (CP) and lactoferrin (LF) play an important role in regulation of inflammation and oxidative stress in vertebrates. It was previously shown that these proteins may work synergetically as antimicrobial and anti-inflammatory agents by forming complexes, such as MPO-CP and LF-CP. However, interaction of metalloprotein molecules with each other has never been characterized at a single-molecule level.

Aggregation of Influenza A Virus Nuclear Export Protein

Influenza A virus nuclear export protein (NEP) plays an important role in the viral life cycle. Recombinant NEP proteins containing (His)₆-tag at either N-or C-terminus were obtained by heterologous expression in Escherichia coli cells and their high propensity for aggregation was demonstrated. Dynamic light scattering technique was used to study the kinetics and properties of NEP aggregation in solutions under different conditions (pH, ionic strength, presence of low-molecular-weight additives and organic solvents). Using atomic force microscopy, the predominance of spherical aggregates in all examined NEP preparations was shown, with some amyloid-like structures being observed in the case of NEP-C protein. A number of structure prediction programs were used to identify aggregation-prone regions in the NEP structure. All-atom molecular dynamics simulations indicate a high rate of NEP molecule aggregation and reveal the regions preferentially involved in the intermolecular contacts that are located at the edges of the rod-like protein molecule. Our results suggest that NEP aggregation is determined by different types of interactions and represents an intrinsic property of the protein that appears to be necessary for its functioning in vivo.     

Study of spectra of detached electrons produced in collisions of negative deuterium ions

Spectra of detached electrons produced in mutual collisions of D⁻ ions in the relative energy range of 1.8−6.1 eV were investigated. In addition to electrons corresponding to conversion of D⁻ ions into D⁰ atoms, peaks corresponding to the production of D₂ molecules and D₂⁻ molecular ions in the ground electronic state were revealed. The existence of D₂⁻ ions over a time longer than the period of molecular oscillations was confirmed experimentally for the first time.     

Structural and functional changes in thrombocytes in experimental atherosclerosis

Electron paramagnetic resonance with the use of stearic acid derivatives (5- and 16-doxylstearate) as spin probes was applied to studies of the structural organization of rabbit platelets in experimental atherosclerosis. Substantial differences were established in the molecular packing of phospholipid plasma membranes, associated with a higher molar content of cholesterol in the cells. An increase in the aggregation properties of platelets was also observed, manifesting in a shorter time of the ADP-induced aggregation of platelets isolated from plasma. The data obtained confirm the primary part of membranotropic cholesterol activity in atherosclerosis, attesting to the validity of the "membraneous" hypothesis of the atherogenesis.     

Larvae of related Diptera species from thermally contrasting habitats exhibit continuous up-regulation of heat shock proteins and high thermotolerance

A population of Stratiomys japonica, a species belonging to the family Stratiomyidae (Diptera), common name ‘soldier flies’, occurs in a hot volcanic spring, which is apparently among the most inhospitable environments for animals because of chemical and thermal conditions. Larvae of this species, which naturally often experience temperatures more than 40 °C, have constitutively high concentrations of the normally inducible heat-shock protein Hsp70, but very low level of corresponding mRNA. Larvae of three other species of the same family, Stratiomys singularior, Nemotelus bipunctatus and Oxycera pardalina, are confined to different type semi-aquatic habitats with contrasting thermal regime. However, all of them shared the same pattern of Hsp70 expression. Interestingly, heat-shock treatment of S. japonica larvae activates heat-shock factor and significantly induces Hsp70 synthesis, whereas larvae of O. pardalina, a species from constant cold environment, produce significantly less Hsp70 in response to heat shock. Adults of the four species also exhibit lower, but detectable levels of Hsp70 without heat shock. Larvae of all species studied have very high tolerance to temperature stress in comparison with other Diptera species investigated, probably representing an inherent adaptive feature of all Stratiomyidae enabling successful colonization of highly variable and extreme habitats.