WoS İndeksli Yayınlar Koleksiyonu / WoS Indexed Publications Collection

Permanent URI for this collectionhttps://hdl.handle.net/11147/7150

Browse

Filters

2024 - 20252

Settings

Author: search.filters.author.Bogdanov, A. M.Language: search.filters.language.en

Search Results

Now showing 1 - 2 of 2
  • Article
    Citation - WoS: 1
    Citation - Scopus: 1
    Two Key Substitutions in the Chromophore Environment of mKate2 Produce an Enhanced FusionRed-Like Red Fluorescent Protein
    (Russian Federation Agency Science & innovation, 2025) Ruchkin, D. A.; Gavrikov, A. S.; Kolesov, D., V; Gorokhovatsky, A. Yu.; Chepurnykh, T., V; Mishin, A. S.; Bogdanov, A. M.
    Red fluorescent proteins (RFPs) are often probes of choice for living tissue microscopy and whole-body imaging. When choosing a specific RFP variant, the priority may be given to the fluorescence brightness, maturation rate, monomericity, excitation/emission wavelengths, and low toxicity, which are rarely combined in an optimal way in a single protein. If additional requirements such as prolonged fluorescence lifetime and/or blinking ability are applied, the available repertoire of probes could dramatically narrow. Since the entire diversity of conventional single-component RFPs belongs to just a few phylogenetic lines (DsRed-, eqFP578-and eqFP611-derived being the major ones), it is not unexpected that their advantageous properties are split between close homologs. In such cases, a systematic mutagenetic analysis focusing on variant-specific amino acid residues can shed light on the origins of the distinctness between related RFPs and may aid in consolidating their strengths in new RFP variants. For instance, the protein FusionRed, despite being efficient in fluorescence labeling thanks to its good monomericity and low cytotoxicity, has undergone considerable loss in fluorescence brightness/lifetime compared to the parental mKate2. In this contribution, we describe a fast-maturing monomeric RFP designed semi-rationally based on the mKate2 and FusionRed templates that outperforms both its parents in terms of molecular brightness, has extended fluorescence lifetime, and displays a spontaneous blinking pattern that is promising for nanoscopy use.
  • Article
    Mkate2-k67r/R197h-extra-bright Red Fluorescent Biomarker of New Generation. X-Ray Structure and Molecular Dynamic Properties
    (Maik Nauka/interperiodica/springer, 2024) Goryacheva, E. A.; Rossokhin, A. V.; Ruchkin, D. A.; Bogdanov, A. M.; Artemyev, I. V.; Pletneva, N. V.; Plenev, V. Z.
    Objective: Cell biology continuously shows the need for new fluorescent tags with advanced properties. The object of our current study is a new genetically encoded monomeric red fluorescent biomarker mKate2-K67R/R197H (lambda ex/lambda em 579/603 mn), designed from commercial biomarker mKate2 by two R197H/K67R mutations. The mKate2 precursor, a far-red fluorescent protein, is nearly 3-fold brighter than the previously designed mKate. Compared with commercial mKate2, the double mutant mKate2-K67R/R197H (alternative names FusionRed2 and Diogenes) exhibits an additional similar to 1.6-fold increase in fluorescence brightness and represents the next generation of extra-bright red fluorescent probes offering novel possibilities for fluorescent imaging of proteins in living cells and animals. Methods: The paper presents the results of X-ray and molecular dynamics study of new bright biomarker mKate2-K67R/R197H. Results and Discussion: The three dimensional structure of new advanced red fluorescent biomarker mKate2-K67R/R197H has been studied by X-ray method at 1.5 angstrom resolution supported by molecular dynamics (MD) study The principal structural fold of the protein is an 11-stranded beta-barrel. The nearest chromophore environment (<= 4 angstrom) comprises 18 tightly packed residues. Conclusions: The MD study showed that the brightness of mKate2-K67R/R197H and its mKate2 precursor correlates with the dipole moments of the amino acid environments of the chromophores. The higher the dipole moment, the higher the brightness of biomarker.