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Ligand-Binding Activity of Recombinant Tetracycline Antibiotics Receptor TetR as Determined by Fluorescence Spectroscopy
Abstract
The ligand-binding activity of recombinant receptor TetR towards tetracycline (Tc) and its analogues (TC) has been studied by fluorescence spectroscopy. The addition of Tc to the functionally active protein in a solution quenched TetR tryptophan fluorescence by 80% and greatly enhanced the emission of Tc. The formation of the TetR-Tc complex was manifested by the appearance of a band in the Tc fluorescence region with a maximum at 510 nm, resulting from non-radiative energy transfer (FRET effect) from an excited tryptophan residue at 280 nm to the ligand. Other TCs (4-epi-Tc and lymecycline) also bound to TetR, exhibiting the properties of fluorescent probes. The spectral effects of complexation were reduced or completely disappeared in cases of partially or completely denatured TetR in the presence of urea. Therefore, fluorescence spectroscopy can serve as a reliable tool for assessing the TC-binding activity of recombinant TetR in the course of its preparation, storage and technological processing for practical use as a key component of bioanalytical systems for the determination of tetracycline antibiotics in food.
About the Authors
T. S. Serchenya
Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus
Belarus
Belarus
Minsk
V. S. Lapina
Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus
Belarus
Belarus
Minsk
O. V. Sviridov
Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus
Belarus
Belarus
Minsk
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Review
For citations:
Serchenya T.S., Lapina V.S., Sviridov O.V. Ligand-Binding Activity of Recombinant Tetracycline Antibiotics Receptor TetR as Determined by Fluorescence Spectroscopy. Zhurnal Prikladnoii Spektroskopii. 2025;92(5):603-611. (In Russ.)
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