Сибирский федеральный университет

Тип публикации: статья из журнала

Год издания: 2021

Идентификатор DOI: 10.3390/ijms221910449

Ключевые слова: bacterial luciferase, conforma-tional stability, fret, molecular dynamics, time-resolved spectroscopy, tryptophan fluorescence, unfolding pathway, urea-induced denaturation

Аннотация: Detecting the folding/unfolding pathways of biological macromolecules is one of the urgent problems of molecular biophysics. The unfolding of bacterial luciferase from Vibrio harveyi is well-studied, unlike that of Photobacterium leiognathi, despite the fact that both of them are actively used as a reporter system. The aim of this study was to compare the conformational transitions of these luciferases from two different protein subfamilies during equilibrium unfolding with urea. Intrinsic steady-state and time-resolved fluorescence spectra and circular dichroism spectra were used to determine the stages of the protein unfolding. Molecular dynamics methods were applied to find the differences in the surroundings of tryptophans in both luciferases. We found that the unfolding pathway is the same for the studied luciferases. However, the results obtained indicate more stable tertiary and secondary structures of P. leiognathi luciferase as compared to enzyme from V. harveyi during the last stage of denaturation, including the unfolding of individual subunits. The distinctions in fluorescence of the two proteins are associated with differences in the structure of the C-terminal domain of α-subunits, which causes different quenching of tryptophan emissions. The time-resolved fluorescence technique proved to be a more effective method for studying protein unfolding than steady-state methods. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Журнал: International Journal of Molecular Sciences

Выпуск журнала: Vol. 22, Is. 19

Номера страниц: 10449

ISSN журнала: 16616596

Издатель: MDPI

• Nemtseva Elena (Siberian Fed Univ, Krasnoyarsk 660041, Russia; Inst Biophys SB RAS, Photobiol Lab, Krasnoyarsk 660036, Russia)
• Gulnov Dmitry (Siberian Fed Univ, Krasnoyarsk 660041, Russia)
• Gerasimova Marina A. (Siberian Fed Univ, Krasnoyarsk 660041, Russia)
• Sukovatyi Lev A. (Siberian Fed Univ, Krasnoyarsk 660041, Russia)
• Burakova Ludmila P. (Siberian Fed Univ, Krasnoyarsk 660041, Russia; Inst Biophys SB RAS, Photobiol Lab, Krasnoyarsk 660036, Russia)
• Karuzina Natalya E. (Siberian Fed Univ, Krasnoyarsk 660041, Russia)
• Melnik Bogdan S. (Russian Acad Sci, Inst Prot Res, Pushchino 142290, Russia)
• Kratasyuk Valentina A. (Siberian Fed Univ, Krasnoyarsk 660041, Russia; Inst Biophys SB RAS, Photobiol Lab, Krasnoyarsk 660036, Russia)

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