Enabling Fluorescence Lifetime Imaging Multiplexing Using UnaG through Its Modification with Canonical and Noncanonical Amino Acids.

Fluorogen-activating proteins are powerful molecular tools for microscopy, including functional imaging. These proteins serve as an alternative to GFP-like proteins, as they do not require oxygen for chromophore maturation. However, the restricted selectivity of proteins to chromophores, combined with the limited number of spectral channels of conventional fluorescent microscopes, hinders the development of multicolor synthetic dyes.

Biochemical characterization of Selaginella 14-3-3 protein isoforms provides insights into the evolution of their phylogenetic groups.

Numerous isoforms of 14-3-3 proteins regulate plant growth, nutrient uptake, flowering, and signaling by interacting with phosphorylated partner proteins. Recently, the two major phylogenetic groups of 14-3-3 isoforms in Arabidopsis thaliana, epsilon and non-epsilon, were found to have distinct biochemical properties, but 14-3-3 proteins in non-model and early-diverging plant lineages remain unexplored. Here, we report a comparative study of 14-3-3 isoforms from Selaginella moellendorffii, a lycophyte representing ancient vascular plants.

Ultra-potent RBM-specific single-domain antibody broadly neutralizes multiple SARS-CoV-2 variants with picomolar activity.

SARS-CoV-2 infection remains a cause of severe illness in high-risk individuals, with few antiviral agents currently available. The emergence of new SARS-CoV-2 variants accumulating an increasing number of mutations significantly challenges the development of effective therapeutics. We describe broadly neutralizing single-domain antibody 1p1B10 having picomolar activity against both previously circulating SARS-CoV-2 variants, including Wuhan D614G, Alpha, Beta, Gamma, Delta and Omicron BA.

Establishment of an efficient one-step enzymatic synthesis of cyclic-2,3-diphosphoglycerate.

Extremolytes - unique compatible solutes produced by extremophiles - protect biological structures like membranes, proteins, and DNA under extreme conditions, including extremes of temperature and osmotic stress. These compounds hold significant potential for applications in pharmaceuticals, healthcare, cosmetics, and life sciences. However, despite their considerable potential, only a limited number of extremolytes - most notably ectoine and hydroxyectoine - have achieved commercial relevance, primarily due to the absence of efficient production strategies for the majority of other extremolytes.

A green dichromophoric protein enabling foliage mimicry in arthropods.

Molecular mechanisms underlying the green insect camouflage have puzzled researchers for over a century. Here, we isolated and identified a green water-soluble protein from the integument of bush-cricket . De novo sequencing and cloning revealed a severely fragmented form of vitellogenins, ubiquitous and multifunctional, but still largely enigmatic glycolipoproteins essential for embryonic development and lacking structural characterization.

High-resolution structure reveals enhanced 14-3-3 binding by a mutant SARS-CoV-2 nucleoprotein variant with improved replicative fitness.

Replication of many viruses depends on phosphorylation of viral proteins by host protein kinases and subsequent recruitment of host protein partners. The nucleoprotein (N) of SARS-CoV-2 is heavily phosphorylated and recruits human phosphopeptide-binding 14-3-3 proteins early in infection, which is reversed prior to nucleocapsid assembly in new virions. Among the multiple phosphosites of N, which are particularly dense in the serine/arginine-rich interdomain region, phospho-Thr205 is highly relevant for 14-3-3 recruitment by SARS-CoV-2 N.

Structural and functional insights into extreme thermal stability and activity of two GH12 domains of a multidomain glycosidase from a hyperthermophilic euryarchaeon.

Bacteria and fungi are well known for efficient degradation of plant polysaccharides thanks to various enzymes involved in plant cell wall decomposition. However, little is known about the role of archaea in this process or the repertoire and features of their polysaccharide-degrading enzymes. In our previous work, we discovered an archaeal multidomain glycosidase (MDG) composed of three catalytic domains (GH5 and two GH12) and two cellulose-binding modules (CBM2).

Mechanisms of Virulence of and Interaction with the Host Immune System.

is a non-tuberculosis fast-growing mycobacterium that has recently become a serious concern due to its rapidly increasing prevalence worldwide, mainly in individuals with a high susceptibility to pulmonary infections, for example, patients with cystic fibrosis, bronchiectasis, chronic obstructive pulmonary disease, and previous tuberculosis infection. According to present estimations, at least 20% of patients with cystic fibrosis are infected with . .

The Big, Mysterious World of Plant 14-3-3 Proteins.

14-3-3 is a family of small regulatory proteins found exclusively in eukaryotic organisms. They selectively bind to phosphorylated molecules of partner proteins and regulate their functions. 14-3-3 proteins were first characterized in the mammalian brain approximately 60 years ago and then found in plants, 30 years later.

Assembly of allophycocyanin from individual subunits: involvement of apo-form in the formation of trimers.

In cyanobacteria and red algae, allophycocyanin (APC), as well as other phycobiliproteins, is involved in the energy transfer of photosystems. Since APC is a potent fluorescent protein for imaging and biomedical applications, it is necessary to obtain purified protein in large quantities, which is currently possible by biosynthesis in bacterial systems. Here we emphasize the challenges of obtaining the trimeric form of the protein from α-APC and β-APC subunits of allophycocyanin in vitro.

Biochemical signatures strongly demarcate phylogenetic groups of plant 14-3-3 isoforms.

Interaction of dimeric 14-3-3 proteins with phosphotargets regulates various physiological processes in plants, from flowering to transpiration and salt tolerance. Several genes express distinct 14-3-3 "isoforms," particularly numerous in plants, but these are unevenly studied even in model species. Here we systematically investigated twelve 14-3-3 isoforms from Arabidopsis thaliana.

Tenuifilum osseticum sp. nov., a novel thermophilic hydrolytic bacterium within the Tenuifilaceae isolated from a North Ossetian thermal spring, and emended description of the genus Tenuifilum.

A novel anaerobic moderately thermophilic bacterium, strain 4138-str, was isolated from a thermal spring of North Ossetia (Russian Federation). Gram-negative cells were non-sporeforming, straight or curved filamentous rods, occasionally forming rosettes. The strain grew at 30-55 °C, pH range of 6.

Mutational interference with oligomerization properties of OCP-related apo- and holoproteins studied by analytical ultracentrifugation.

In this study, the oligomerization pattern of apo- and holoforms of the Orange Carotenoid Protein (OCP) was examined under different conditions such as photoactivation state, concentration, and carotenoid embedment using analytical ultracentrifugation. Furthermore, studies were conducted on OCP constructs carrying point mutations of amino acid residues affecting OCP oligomerization. Our findings reveal that the concentration-dependent dimerization of dark-adapted OCP holoprotein from Synechocystis sp.

Anaeroselena agilis gen. nov., sp. nov., a Novel Sulfite- and Arsenate-Respiring Bacterium Within the Family Acetonemataceae Isolated from a Thermal Spring of North Ossetia.

A novel Gram-negative, motile, rod-shaped bacterium, designated 4137-cl, was isolated from a thermal spring of North Ossetia (Russian Federation). Strain 4137-cl grew at 30-50 °C (optimum 42 °C) with 0-3.5% NaCl (optimum 0-0.

Enhancing aerobic composting of food waste by adding hydrolytically active microorganisms.

The biomass of native microorganisms in food waste (FW) suitable for accelerated composting is initially low and requires time for adaptation. Adding of efficient hydrolytic microorganisms should be able to enhance compost-specific microbial activity, adjust microbial community structure, and potentially hasten FW biodegradation. This study aimed to identify bacterial and fungal strains with growth characteristics suitable for accelerating FW composting.

sp. nov., a Thermophilic, Obligately Chemolithoautotrophic, Sulfur-Oxidizing Bacterium from a Hot Spring and Proposal of fam. nov.

An aerobic, obligately chemolithoautotrophic, sulfur-oxidizing bacterium, strain AK1, was isolated from a terrestrial hot spring of the Uzon Caldera, Kamchatka, Russia. The cells of the new isolate were Gram-negative motile rods with a single polar flagellum. Strain AK1 grew at 37-55 °C (optimum 50 °C) with 0-1.

Antioxidant properties of the soluble carotenoprotein AstaP and its feasibility for retinal protection against oxidative stress.

Photodamage to the outer segments of photoreceptor cells and their impaired utilization by retinal pigment epithelium (RPE) cells contribute to the development of age-related macular degeneration (AMD) leading to blindness. Degeneration of photoreceptor cells and RPE cells is triggered by reactive oxygen species (ROS) produced by photochemical reactions involving bisretinoids, by-products of the visual cycle, which accumulate in photoreceptor discs and lipofuscin granules of RPE. Carotenoids, natural antioxidants with high potential efficacy against a wide range of ROS, may protect against the cytotoxic properties of lipofuscin.

Removal of terpenes in the presence of easily degradable compounds during biofiltration of gas emissions from composting of municipal solid waste.

Composting of the organic fraction of municipal solid waste (OFMSW) is accompanied by the emission of large volumes of harmful, hazardous and foul-smelling volatile organic compounds (VOCs). To improve the efficiency of terpenes removal, which constitute a significant part of VOCs, pure cultures of microorganisms dominating in its microbiota were isolated from the microbial community of the biofilter, which has been cleaning such emissions for a long time. Seven pure cultures were isolated and then tested for being able to grow on a mineral medium in the presence of terpene vapor as the only source of carbon and energy.

Re-engineering of a carotenoid-binding protein based on NMR structure.

Recently, a number of message passing neural network (MPNN)-based methods have been introduced that, based on backbone atom coordinates, efficiently recover native amino acid sequences of proteins and predict modifications that result in better expressing, more soluble, and stable variants. However, usually, X-ray structures, or artificial structures generated by algorithms trained on X-ray structures, were employed to define target backbone conformations. Here, we show that commonly used algorithms ProteinMPNN and SolubleMPNN display low sequence recovery on structures determined using NMR.

Link Between Individual Codon Frequencies and Protein Expression: Going Beyond Codon Adaptation Index.

An important role of a particular synonymous codon composition of a gene in its expression level is well known. There are a number of algorithms optimizing codon usage of recombinant genes to maximize their expression in host cells. Nevertheless, the underlying mechanism remains unsolved and is of significant relevance.

luxA Gene From Enhygromyxa salina Encodes a Functional Homodimeric Luciferase.

Several clades of luminescent bacteria are known currently. They all contain similar lux operons, which include the genes luxA and luxB encoding a heterodimeric luciferase. The aldehyde oxygenation reaction is presumed to be catalyzed primarily by the subunit LuxA, whereas LuxB is required for efficiency and stability of the complex.

Degradation of high concentrations of commercial polylactide packaging on food waste composting in pilot-scale test.

The increasing use of synthetic biodegradable polymers, such as aliphatic polyesters, has led to a greater need to understand their behavior in an end-of-life scenario as food packaging materials. The aim of this work was to investigate the effect on composting of high to 10 wt% concentration of commercial polylactide packaging in food waste during a 98-day pilot-scale test. Members of the genera Bacillus, Geobacillus, Caldibacillus, Compostibacillus, Novibacillus, Planifilum and Aeribacillus accounted for 77 % of the bacterial community at the initial stage.

Effects of Nitrosyl Iron Complexes with Thiol, Phosphate, and Thiosulfate Ligands on Hemoglobin.

Nitrosyl iron complexes are remarkably multifactorial pharmacological agents. These compounds have been proven to be particularly effective in treating cardiovascular and oncological diseases. We evaluated and compared the antioxidant activity of tetranitrosyl iron complexes (TNICs) with thiosulfate ligands and dinitrosyl iron complexes (DNICs) with glutathione (DNIC-GS) or phosphate (DNIC-PO) ligands in hemoglobin-containing systems.