Comparison of the Structural Dynamics of Bacterial Luciferases Using Time-Resolved Luminescence Techniques

Описание

Тип публикации: статья из журнала

Год издания: 2024

Идентификатор DOI: 10.1134/S1062873824709310

Ключевые слова: phosphorescence, fluorescence, Dipolar relaxation, tryptophan, protein dynamics, bioluminescence, luciferase

Аннотация: The structural dynamics of two proteins—luciferases from bacteria <i>Vibrio harveyi</i> and <i>Photobacterium leiognathi</i>, was studied using time-resolved fluorescence and phosphorescence of the tryptophan residues at 5–35°C. Earlier, the temperature effects on functional properties of these luciferases were found to be different. Revealing the structural and dynamic basis of the difference will contribute to understanding the principles of molecular adaptation of cells for living in unfavorable conditions. Nanosecond-resolved shifts of the fluorescence spectra were analyzed and dipolar relaxation times of the tryptophans microenvironment were obtained: τR1 = 0.22–0.49 ns and τR2 = 8.04–9.37 ns for <i>P. leiognathi</i> luciferase and τR1 = 0.18–0.27 ns and τR2 = 6.3–9.0 ns for <i>V. harveyi</i> luciferase. Temperature dependences of τR1 indicate a more rigid structure of the latter protein. Microsecond-resolved phosphorescence decays were measured and phosphorescence lifetimes τp1 and τp2 were determined: 0.15–0.30 and 0.91–2.77 ms for <i>P. leiognathi</i> luciferase and 0.16–0.33 and 2.27–5.45 ms for <i>V. harveyi</i> luciferase, respectively. The increased values of τp2 for <i>V. harveyi</i> luciferase also indicate more rigid microenvironment of the tryptophans within this protein. The experimental data were compared with the molecular modeling results on the mobility of tryptophans and adjacent residues in bacterial luciferase structures under temperature variations.

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Издание

Журнал: Bulletin of the Russian Academy of Sciences: Physics

Выпуск журнала: Т. 88, № S2

Номера страниц: 217-223

ISSN журнала: 10628738

Место издания: Moscow

Издатель: Pleiades Publishing, Ltd.

Персоны

  • Nemtseva E.V. (Institute of Biophysics, Siberian Branch, Russian Academy of Sciences)
  • Gulnov D.V. (Siberian Federal University)
  • Eroshenko P.A. (Siberian Federal University)
  • Sukovatyi L.A. (Orbeli Institute of Physiology, National Academy of Sciences of the Republic of Armenia)
  • Kratasyuk V.A. (Institute of Biophysics, Siberian Branch, Russian Academy of Sciences)

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