292 research outputs found
«В очікуванні залізниці»: Як периферія сполучалася з центрами індустріалізації
3-5 листопада 2011 р. у м. Львові відбулася Міжнародна конференція з залізничної історії «В очікуванні залізниці»: Як периферія сполучалася з центрами індустріалізації», присвячена 150-літтю відкриття першого залізничної лінії на території сучасної України
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Kinetic analysis of protein aggregation monitored by real-time 2D solid-state NMR spectroscopy
It is shown that real-time 2D solid-state NMR can be used to obtain kinetic and structural information about the process of protein aggregation. In addition to the incorporation of kinetic information involving intermediate states, this approach can offer atom-specific resolution for all detectable species. The analysis was carried out using experimental data obtained during aggregation of the 10.4 kDa Crh protein, which has been shown to involve a partially unfolded intermediate state prior to aggregation. Based on a single real-time 2D 13C–13C transition spectrum, kinetic information about the refolding and aggregation step could be extracted. In addition, structural rearrangements associated with refolding are estimated and several different aggregation scenarios were compared to the experimental data
Biological solid-state NMR: Integrative across different scientific disciplines
For almost five decades, solid-state NMR (ssNMR) has been used to study complex biomolecular systems. This article gives a view on how ssNMR methods and applications have evolved during this time period in a broader structural biology context. It also discusses possible directions for additional developments and the future role of ssNMR in a life science context and beyond
Роль хламидийной инфекции в патологии человека
Освещены биологические свойства хламидий и вопросы эпидемиологии хламидийной инфекции. Показана ее большая распространенность, в том числе в Украине, и существенное патогенное влияние на здоровье мужчин, женщин и детей.Biological properties of Chlamydia and the problems of epidemiology of Chlamydia infection are featured. Its high frequency in many countries including Ukraine and considerable pathogenic influence on the health of men, women, and children are shown
In vivo Assembly of Artificial Metalloenzymes and Application in Whole-Cell Biocatalysis
We report the supramolecular assembly of artificial metalloenzymes (ArMs), based on the Lactococcal multidrug resistance regulator (LmrR) and an exogeneous copper(II)-phenanthroline complex, in the cytoplasm of E. coli cells. A combination of catalysis, cell-fractionation and inhibitor experiments, supplemented with in-cell solid-state NMR, confirmed the in-cell assembly. The ArM containing whole cells were active in the catalysis of the enantioselective Friedel-Crafts alkylation of indoles and the Diels-Alder reaction of azachalcone with cyclopentadiene. Directed evolution resulted in two different improved mutants for both reactions, LmrR_A92E_M8D and LmrR_A92E_V15A, respectively. The whole-cell ArM system requires no engineering of the microbial host, the protein scaffold or the cofactor to achieve ArM assembly and catalysis. We consider this a key step towards integrating abiological catalysis in biosynthesis, achieving a hybrid metabolism
Divide and Conquer: A Tailored Solid-state NMR Approach to Study Large Membrane Protein Complexes
Membrane proteins are known to exert many essential biological functions by forming complexes in cell membranes. An example refers to the β-barrel assembly machinery (BAM), a 200 kDa pentameric complex containing BAM proteins A–E that catalyzes the essential process of protein insertion into the outer membrane of gram-negative bacteria. While progress has been made in capturing three-dimensional structural snapshots of the BAM complex, the role of the lipoprotein BamC in the complex assembly in functional lipid bilayers has remained unclear. We have devised a component-selective preparation scheme to directly study BamC as part of the entire BAM complex in lipid bilayers. Combination with proton-detected solid-state NMR methods allowed us to probe the structure, dynamics, and supramolecular topology of full-length BamC embedded in the entire complex in lipid bilayers. Our approach may help decipher how individual proteins contribute to the dynamic formation and functioning of membrane protein complexes in membranes
Бесплатный обязательный экземпляр полиграфических и других изданий как один из источников комплектования Латвийской академической библиотеки
Aggregation of amyloid beta (Aβ) into oligomers and fibrils is believed to play an important role in the development of Alzheimer's disease (AD). To gain further insight into the principles of aggregation, we have investigated the induction of β-sheet secondary conformation from disordered native peptide sequences through lipidation, in 1-2% hexafluoroisopropanol (HFIP) in phosphate buffered saline (PBS). Several parameters, such as type and number of lipid chains, peptide sequence, peptide length and net charge, were explored keeping the ratio peptide/HFIP constant. The resulting lipoconjugates were characterized by several physico-chemical techniques: Circular Dichroism (CD), Attenuated Total Reflection InfraRed (ATR-IR), Thioflavin T (ThT) fluorescence, Dynamic Light Scattering (DLS), solid-state Nuclear Magnetic Resonance (ssNMR) spectroscopy and Electron Microscopy (EM). Our data demonstrate the generation of β-sheet aggregates from numerous unstructured peptides under physiological pH, independent of the amino acid sequence. The amphiphilicity pattern and hydrophobicity of the scaffold were found to be key factors for their assembly into amyloid-like structures
Simultaneous use of solution, solid-state NMR and X-ray crystallography to study the conformational landscape of the Crh protein during oligomerization and crystallization
We explore, using the Crh protein dimer as a model, how information from solution NMR, solid-state NMR and X-ray crystallography can be combined using structural bioinformatics methods, in order to get insights into the transition from solution to crystal. Using solid-state NMR chemical shifts, we filtered intra-monomer NMR distance restraints in order to keep only the restraints valid in the solid state. These filtered restraints were added to solid-state NMR restraints recorded on the dimer state to sample the conformational landscape explored during the oligomerization process. The use of non-crystallographic symmetries then permitted the extraction of converged conformers subsets. Ensembles of NMR and crystallographic conformers calculated independently display similar variability in monomer orientation, which supports a funnel shape for the conformational space explored during the solution-crystal transition. Insights into alternative conformations possibly sampled during oligomerization were obtained by analyzing the relative orientation of the two monomers, according to the restraint precision. Molecular dynamics simulations of Crh confirmed the tendencies observed in NMR conformers, as a paradoxical increase of the distance between the two β1a strands, when the structure gets closer to the crystallographic structure, and the role of water bridges in this context
Binding of micro-nutrients to the cell wall of the fungus Schizophyllum commune
The cell wall fulfils several functions in the biology of fungi. For instance, it provides mechanical strength, interacts with the (a)biotic environment, and acts as a molecular sieve. Recently, it was shown that proteins and β-glucans in the cell wall of Schizophyllum commune bind Cu2+. We here show that the cell wall of this mushroom forming fungus also binds other (micro-)nutrients. Ca2+, Mg2+, Mn2+, NO3–, PO43-, and SO42- bound at levels > 1 mg per gram dry weight cell wall, while binding of BO3-, Cu2+, Zn2+ and MoO42- was lower. Sorption of Ca2+, Mn2+, Zn2+ and PO43- was promoted at alkaline pH. These compounds as well as BO33-, Cu2+, Mg2+, NO3–, and SO42- that had bound at pH 4, 6, or 8 could be released from the cell wall at pH 4 with a maximum efficiency of 46–93 %. Solid-state NMR spectroscopy showed that the metals had the same binding sites as Cu2+ when a low concentration of this ion is used. Moreover, data indicate that anions bind to the cell wall as well as to the metal ions. Together, it is shown that the cell wall of S. commune binds various (micro-)nutrients and that this binding is higher than the uptake by hyphae. The binding to the cell wall may be used as a storage mechanism or may reduce availability of these molecules to competitors or prevent toxic influx in the cytoplasm
Catalytic fast pyrolysis of biomass : catalyst characterization reveals the feed-dependent deactivation of a technical ZSM-5-based catalyst
Catalyst deactivation due to coking is a major challenge in the catalytic fast pyrolysis (CFP) of biomass. Here, a multitechnique investigation of a technical Al2O3-bound ZSM-5-based extrudate catalyst, used for the CFP of pine wood and cellulose (at a reactor temperature of 500 °C), provided insight into the effects of extrusion, the catalytic pyrolysis process, and catalyst regeneration on the catalyst structure. As a result of a reduction in acidity and surface area due to the coking catalyst, the activity dropped drastically with increasing time-on-stream (TOS), as evidenced by a decrease in aromatics yield. Strikingly, confocal fluorescence microscopy at the single-particle level revealed that vapor components derived from whole biomass or just the cellulose component coke differently. While pine-wood-derived species mainly blocked the external area of the catalyst particle, larger carbon deposits were formed inside the catalyst’s micropores with cellulose-derived species. Pyridine FT-IR and solid-state NMR spectroscopy demonstrated irreversible changes after regeneration, likely due to partial dealumination. Taken together with <30 g kg–1 aromatics yield on a feed basis, the results show a mismatch between biomass pyrolysis vapors and the technical catalyst used due to a complex interplay of mass transfer limitations and CFP chemistry
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