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<p>The ligand-binding activity of the recombinant receptor TetR towards tetracycline (Tc) and its analogs (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 indicated by the appearance of a band in the tetracycline fluorescence region with a maximum at 510 nm resulting from nonradiative energy transfer (FRET effect) from a tryptophan residue excited at 280 nm to the ligand. Other tetracyclines (TC) (4-epi-TC and lymecycline) also bind to TetR, exhibiting the properties of fluorescent probes. The spectral effects of complexation were reduced or even completely eliminated in cases of partially or completely denatured TetR in the presence of urea. Thus, fluorescence spectroscopy can serve as a reliable tool for assessing the TC-binding activity of recombinant TetR during its preparation, storage, and processing for practical use as a key component of bioanalytical systems for determining traces of tetracycline antibiotics in food.</p>

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Fluorescence Spectroscopy Study of the Ligand-Binding Activity of the Recombinant Tetracycline Antibiotics Receptor TetR

  • 期刊论文

  • 发表日期：

    2025年11月4日

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  • T. S. Serchenya,
  • V. S. Lapina,
  • Ο.V. Sviridov

摘要

The ligand-binding activity of the recombinant receptor TetR towards tetracycline (Tc) and its analogs (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 indicated by the appearance of a band in the tetracycline fluorescence region with a maximum at 510 nm resulting from nonradiative energy transfer (FRET effect) from a tryptophan residue excited at 280 nm to the ligand. Other tetracyclines (TC) (4-epi-TC and lymecycline) also bind to TetR, exhibiting the properties of fluorescent probes. The spectral effects of complexation were reduced or even completely eliminated in cases of partially or completely denatured TetR in the presence of urea. Thus, fluorescence spectroscopy can serve as a reliable tool for assessing the TC-binding activity of recombinant TetR during its preparation, storage, and processing for practical use as a key component of bioanalytical systems for determining traces of tetracycline antibiotics in food.