Fluorescent Tags in Biology: Three-Dimensional Structure
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EW ARTICLE
Fluorescent Tags in Biology: Three-Dimensional Structure N. V. Pletnevaa, E. A. Goryachevaa, I. V. Artemyeva, S. F. Arkhipovaa, and V. Z. Pletneva, 1 aShemyakin–Ovchinnikov
Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997 Russia
Received January 28, 2020; revised February 13, 2020; accepted February 18, 2020
Abstract—Fluorescent tags of various natures are widely used in cell biology for visualization and study processes in living organisms including gene expression, localization and migration of proteins and cells of interest, determination of the vital intracellular characteristics such as pH, ion concentration, temperature, etc. The review presents an overview of the three dimensional organization as well as advantages and disadvantages of the most promising fluorescent molecular instruments that have been widely used in biology. Keywords: 3D structure, fluorescent tags, biomarkers low-molecular-weight fluorophores, GFP-like proteins, photoreceptors, phytochromes, flavoproteins, lipocalins DOI: 10.1134/S1068162020040160
INTRODUCTION The rapid development of directions for the creation of fluorescent instruments of both low molecular weight and protein nature is dictated by the urgent needs of biology and biomedicine for studying processes in living organisms. Significant progress in this area has been recently marked by two Nobel Prizes in chemistry: 2008—for the discovery and use of green fluorescent protein GFP; 2014—for the creation of ultra-high resolution fluorescence microscopy. Currently, the creation of fluorescent labels for biology is developing in four directions: (1) organic and inorganic low-molecular-weight fluorophores, (2) GFPlike proteins with a chromophore formed as part of the polypeptide chain as a result of posttranslational modification, (3) photoreceptors (phytochrome and flavoprotein domains), autocatalytically binding the natural chromophores present in the cell, (4) fluorogenactivating proteins (antibodies and lipocalins ), forming noncovalent specific complexes with organic lowmolecular-weight fluorogens of various natures. The creation of new fluorescence constructs based on protein platforms includes a rational design based on sitedirected mutagenesis and directed evolution based on optimizing properties by random mutagenesis. I. ORGANIC AND INORGANIC LOWMOLECULAR-WEIGHT FLUOROPHORES Fluorescent organic compounds, represented by a large group of small molecules, form an ensemble of effective tools for visualizing processes in living cells and organisms. Since the first synthesis of fluorescein in 1871, organic low-molecular-weight fluorophores 1 Corresponding author: e-mail: [email protected].
have been successfully used in cell biology [1–4]. Individual fluorophores have found application for stoichiometric labeling of almost any protein, even in living cells. Other fluorophores have the property of specific binding to DNA or biological membranes. Currently, the variety of properties of such fluorescent labels allows us to determine met
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