A5042580739- Advanced Fluorescence Microscopy Techniques
- Photoreceptor and optogenetics research
- bioluminescence and chemiluminescence research
- Click Chemistry and Applications
- Cell Image Analysis Techniques
- Photosynthetic Processes and Mechanisms
- Advanced Electron Microscopy Techniques and Applications
- Nuclear physics research studies
- Advanced Biosensing Techniques and Applications
- Advanced biosensing and bioanalysis techniques
- Evolution and Genetic Dynamics
- Nuclear Physics and Applications
- Luminescence and Fluorescent Materials
- Atomic and Molecular Physics
- Advanced Chemical Physics Studies
- CRISPR and Genetic Engineering
- Biotin and Related Studies
- Estrogen and related hormone effects
- Cancer Research and Treatments
- Photodynamic Therapy Research Studies
- Advanced NMR Techniques and Applications
- Advanced Proteomics Techniques and Applications
- Fluorine in Organic Chemistry
- Protein Structure and Dynamics
- Plant and animal studies
Institute of Bioorganic Chemistry
2015-2024
Planta
2019-2024
Russian Academy of Sciences
2008-2024
Development Agency of Serbia
2024
Institute of Bioorganic Chemistry
2016-2024
V. Peskov Voronezhsky State Nature Biosphere Reserve
2024
Kurchatov Institute
2023
Westlake University
2023
Shanghai Advanced Research Institute
2023
Moscow Institute of Physics and Technology
2023
Bioluminescence is found across the entire tree of life, conferring a spectacular set visually oriented functions from attracting mates to scaring off predators. Half dozen different luciferins, molecules that emit light when enzymatically oxidized, are known. However, just one biochemical pathway for luciferin biosynthesis has been described in full, which only bacteria. Here, we report identification fungal luciferase and three other key enzymes together form biosynthetic cycle caffeic...
Characterizing the fitness landscape, a representation of for large set genotypes, is key to understanding how genetic information interpreted create functional organisms. Here we determined evolutionarily-relevant segment landscape His3, gene coding an enzyme in histidine synthesis pathway, focusing on combinations amino acid states found at orthologous sites extant species. Just 15% acids yeast His3 orthologues were always neutral while impact remaining 85% depended background....
Abstract Liquid–liquid phase separation (LLPS) contributes to the spatial and functional segregation of molecular processes within cell nucleus. However, role played by LLPS in chromatin folding living cells remains unclear. Here, using stochastic optical reconstruction microscopy (STORM) Hi-C techniques, we studied effects 1,6-hexanediol (1,6-HD)-mediated disruption/modulation on higher-order organization cells. We found that 1,6-HD treatment caused enlargement nucleosome clutches their...
Studies of protein fitness landscapes reveal biophysical constraints guiding evolution and empower prediction functional proteins. However, generalisation these findings is limited due to scarceness systematic data on proteins with a defined evolutionary relationship. We characterized the peaks four orthologous fluorescent broad range sequence divergence. While two studied were sharp, other considerably flatter, being almost entirely free epistatic interactions. Mutationally robust proteins,...
Abstract The discovery of the bioluminescence pathway in fungus Neonothopanus nambi enabled engineering eukaryotes with self-sustained luminescence. However, brightness luminescence heterologous hosts was limited by performance native fungal enzymes. Here we report optimized versions that enhance one to two orders magnitude plant, and mammalian hosts, enable longitudinal video-rate imaging.
Abstract A novel class of fluorescent dyes based on conformationally locked GFP chromophore is reported. These are characterized by red‐shifted spectra, high fluorescence quantum yields and pH‐independence in physiological pH range. The intra‐ intermolecular mechanisms radiationless deactivation ABDI‐BF2 fluorophore selective structural locking various conformational degrees freedom were studied. unique combination solvatochromic lipophilic properties together with “infinite” photostability...
A novel method of protein labeling uses the highly dynamic reversible association a cell-permeable fluorogenic dye and lipocalin Blc mutants.
Genetically encoded photosensitizers, proteins that produce reactive oxygen species when illuminated with visible light, are increasingly used as optogenetic tools. Their applications range from ablation of specific cell populations to precise optical inactivation cellular proteins. Here, we report an orange mutant red fluorescent protein KillerRed becomes toxic blue or green light. This new protein, KillerOrange, carries a tryptophan-based chromophore is novel for photosensitizers. We show...
A genetically encoded fluorescent tag for live cell microscopy is presented. This composed of previously published fluorogen-activating protein FAST and a novel fluorogenic derivative green (GFP)-like chromophore with red fluorescence. The reversible binding the fluorogen accompanied by three orders magnitude increase in fluorescence (580-650 nm). proposed dye instantly stains target cellular proteins fused FAST, washes out minute timescale, exhibits higher photostability signal confocal...
Green fluorescent protein (GFP) from a jellyfish, Aequorea victoria, and its mutants are widely used in biomedical studies as markers. In spite of the enormous efforts academia industry toward generating red mutants, no GFP variants with emission maximum at more than 529 nm have been developed during 15 years since cloning. Here, we new strategy molecular evolution aimed red-emitting mutant GFP. As result, succeeded producing first that substantially matures to state excitation maxima 555...
Of the various super-resolution techniques, stimulated emission depletion (STED) microscopy achieves best temporal resolution at high spatial resolution, enabling live-cell imaging beyond diffraction limit. However, STED and most other methods utilize a particular type of information extractable from raw data, namely positions fluorophores. To expand on use we report here dynamic biosensor. Using fluorescent H2O2 sensor HyPer2 for subdiffraction imaging, were able not only to image filaments...
One of the essential characteristics any tag used in bioscience and medical applications is its size. The larger label, more it may affect studied object, distort behavior. In this paper, using NMR spectroscopy X-ray crystallography, we have structure fluorogen-activating protein FAST both apo form complex with fluorogen. We showed that significant change occurs upon interaction ligand. While completely ordered complex, characterized by higher mobility disordering N-terminus. structural...
Jasmonic and salicylic acids are the major hormones involved in plant response to pests pathogens. Here, we engineered autoluminescent plants that report activity of these with up 53-fold contrast. Using consumer-grade cameras, imaged reporter Arabidopsis thaliana Nicotiana benthamiana throughout normal development, attacks
Fluorescence Resonance Energy Transfer (FRET) using fluorescent protein variants is widely used to study biochemical processes in living cells. FRET detection by fluorescence lifetime measurements the most direct and robust method measure FRET. The traditional cyan-yellow based pairs are getting replaced green-red variants. pair enables excitation at a longer wavelength which reduces cellular autofluorescence phototoxicity while monitoring Despite advances sensors, low efficiency dynamic...
Fluorogenic chromophore–protein pairs were predicted by molecular docking and validated experimentally.
Hispidin is a polyketide found in plants and fungi. In bioluminescent fungi, hispidin serves as precursor of luciferin produced by synthases. Previous studies revealed that synthases differ orthologous from non-bioluminescent fungi the absence two domains with predicted ketoreductase dehydratase activities. Here, we investigated hypothesis loss these evolution led to production emergence bioluminescence. We cloned three well their truncated variants, assessed ability produce bioluminescence...