A5003265757- Advanced Fluorescence Microscopy Techniques
- Epigenetics and DNA Methylation
- RNA modifications and cancer
- DNA and Nucleic Acid Chemistry
- RNA and protein synthesis mechanisms
- Photoreceptor and optogenetics research
- RNA Research and Splicing
- Photosynthetic Processes and Mechanisms
- Click Chemistry and Applications
- Biotin and Related Studies
- Carcinogens and Genotoxicity Assessment
- Force Microscopy Techniques and Applications
- Cellular transport and secretion
- DNA Repair Mechanisms
- Receptor Mechanisms and Signaling
- Cancer-related gene regulation
- Chemical Reactions and Isotopes
- CRISPR and Genetic Engineering
- bioluminescence and chemiluminescence research
- Light effects on plants
- Advanced Electron Microscopy Techniques and Applications
- Genomics and Chromatin Dynamics
- Lipid Membrane Structure and Behavior
- Electrochemical Analysis and Applications
- Photodynamic Therapy Research Studies
Kurchatov Institute
2019-2025
Albert Einstein College of Medicine
2010-2013
Lomonosov Moscow State University
2002-2010
New York University
2006-2010
Yeshiva University
2007
Stony Brook University
2006
State University of New York
2006
Abstract Recent advancements in genetically encoded calcium indicators, particularly those based on green fluorescent proteins, have optimized their performance for monitoring neuronal activities a variety of model organisms. However, progress developing red-shifted GECIs, despite advantages over has been slower, resulting fewer options end-users. In this study, we explored topological inversion and soma-targeting strategies, which are complementary to conventional mutagenesis, re-engineer...
A variety of genetically encoded calcium indicators are currently available for visualization dynamics in cultured cells and vivo. Only one them, called NIR-GECO1, exhibits fluorescence the near-infrared region spectrum. NIR-GECO1 is engineered based on fluorescent protein mIFP derived from bacterial phytochromes. However, has an inverted response to ions its excitation spectrum not optimal commonly used 640 nm lasers. Using small phytochrome GAF-FP calmodulin/M13-peptide pair, we developed...
Genetically encoded calcium indicators based on truncated troponin C are attractive probes for imaging due to their relatively small molecular size and twofold reduced ion buffering. However, the best-suited members of this family, YTnC cNTnC, suffer from low brightness, limited dynamic range, and/or poor sensitivity transients in neurons. To overcome these limitations, we developed an enhanced version YTnC, named YTnC2. Compared with YTnC2 had 5.7-fold higher brightness 6.4-fold increased...
DNA damage caused by the binding of tumorigen 7R,8S-diol 9S,10R-epoxide (B[a]PDE), a metabolite bezo[a]pyrene, to guanine in CpG dinucleotide sequences could affect methylation and, thus, represent potential epigenetic mechanism chemical carcinogenesis. In this work, we investigated impact stereoisomeric (+)- and (−)-trans-anti-B[a]P-N2-dG adducts (B+ B-) on prokaryotic methyltransferases M.SssI M.HhaI. These two recognize GCGC sequences, respectively, transfer methyl group C5 atom cytosine...
Eco RII DNA methyltransferase (M.EcoRII) recognizes the 5′…CC*T/AGG…3′ sequence and catalyzes transfer of methyl group from S ‐adenosyl‐ l ‐methionine to C5 position inner cytosine residue (C*). Here, we study mechanism inhibition M.EcoRII by containing 2‐pyrimidinone, a analogue lacking an NH 2 at C4 pyrimidine ring. Also, 2‐pyrimidinone was used for probing contacts with functional groups bases recognition sequence. 2‐Pyrimidinone incorporated into 5′…CCT/AGG…3′ replacing target nontarget...
The biologically most significant genotoxic metabolite of the environmental pollutant benzo[ a ]pyrene (B[ ]P), (+)‐7 R ,8 S ‐diol 9 ,10 ‐epoxide, reacts chemically with guanine in DNA, resulting predominant formation (+)‐ trans ‐B[ ]P‐ N 2 ‐dG and, to lesser extent, cis adducts. Here, we compare effects adduct stereochemistry and conformation on methylation cytosine catalyzed by two purified prokaryotic DNA methyltransferases (MTases), SssI HhaI, lesions positioned within or adjacent their...
NTnC-like green fluorescent genetically encoded calcium indicators (GECIs) with two ion binding sites were constructed using the insertion of truncated troponin C (TnC) from Opsanus tau into proteins (GFPs). These GECIs are small containing N- and C-termini GFP; they exert a limited effect on cellular free concentration; in contrast to calmodulin-based lack undesired interactions intracellular neurons. The available TnC-based NTnC or YTnC had either an inverted response high brightness but...
The PSmOrange and PSmOrange2 fluorescent proteins undergo irreversible photoconversion from the orange to far-red form under non-cytotoxic blue light. Both required high-power light presence of endogenous or exogenous oxidants for efficient photoswitching. Here, we report next-generation version PSmOrange2, called PSmOrange3, which can be efficiently photoconverted (Ex/Em at 550 nm/564 nm) 614 nm/655 with 430-470 nm violet-blue moderate power (100-180 mW/mm2) in a native cellular...
Branched-chain amino acids (BCAAs) play an important role in the functioning of mammalian cells and central nervous system. However, available genetically encoded indicators for BCAAs are based on Förster resonance energy transfer have a limited dynamic range. We developed single fluorescent protein-based sensor BCAAs, called NeIle, which is composed circularly permutated mNeonGreen protein inserted into leucine-isoleucine-valine binding (LIVBP) from
Abstract EcoRII DNA methyltransferase (M.EcoRII) recognizes the sequence 5′…CC*T/AGG…3′ and catalyzes transfer of methyl group from S-adenosyl-L-methionine to C5 position inner cytosine residue (C*). We obtained several duplexes containing photoactive 5-iodo-2′-deoxyuridine (i5dU) or 5-[4-(3-(trifluoromethyl)-3H-diazirin-3-yl)phenyl]-2′-deoxyuridine (Tfmdp-dU) characterize regions M.EcoRII involved in binding investigate double helix conformational changes that take place during methylation....
True genetically encoded monomeric fluorescent timers (tFTs) change their color as a result of the complete transition blue form into red over time. Tandem FTs (tdFTs) consequence fast and slow independent maturation two forms with different colors. However, tFTs are limited to derivatives mCherry mRuby proteins have low brightness photostability. The number tdFTs is also limited, there no blue-to-red or green-to-far-red tdFTs. not previously been directly compared. Here, we engineered novel...
The mRubyFT is a monomeric genetically encoded fluorescent timer based on the mRuby2 protein, which characterized by complete maturation of blue form with subsequent conversion to red one. It has higher brightness in mammalian cells and photostability compared other timers. A high-resolution structure known characteristic chromophore, but structural details its remain obscure. In order obtain insight into this, we obtained an S148I variant (mRubyFTS148I) blocked over time chromophore. X-ray...
Abstract Review: ca. 150 refs.