zhpsЖурнал прикладной спектроскопииZhurnal Prikladnoii Spektroskopii0514-7506B. I. Stepanov Institute of Physics of the National Academy of Scienceszhps-953Research ArticleСтатьиIn vitro и in siliko oценка взаимодействия сывороточного альбумина человека с омега-3-6-9 жирными кислотамиIn vitro and in silico assessment of human serum albumin interactions with omega 3-6-9 fatty acidsValojerdiF. M.ValojerdiF. M.

Тегеран

Tehran

FarasatA.FarasatA.

Казвин

Qazvin

a.farasat@qums.ac.irGheibiN.GheibiN.

Казвин

Qazvin

ngheibi@qums.ac.irИсламский университет АзадDepartment of Biology, Science and Research Branch, Islamic Azad UniversityНаучно-исследовательский институт профилактики неинфекционных заболеваний, Казвинский университет медицинских наукCellular and Molecular Research Center, Research Institute for Prevention of Non-Communicable Diseases, Qazvin University of Medical Sciences202130112021886977977Copyright © Valojerdi F.M., Farasat A., Gheibi N., 20212021Valojerdi F.M., Farasat A., Gheibi N.Valojerdi F.M., Farasat A., Gheibi N.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://zhps.ejournal.by/jour/article/view/953

Взаимодействие между сывороточным альбумином человека (САЧ) и омега-3, омега-6 и омега-9 жирными кислотами как ненасыщенными жирными кислотами исследовано методами спектрофотометрии в УФ-видимом диапазоне, спектроскопии кругового дихроизма (КД), иммуноферментного анализа (ELISA), измерения времени жизни и анизотропии флуоресценции, а также моделирования визуальной молекулярной динамики (МД). Термодинамические параметры термической и химической денатурации САЧ оценивали в присутствии омега-3-6-9 жирных кислот и без них. Для чистого САЧ (298 K) Tm = 327.7 K и ΔG0 = 88 кДж/моль, при обработке САЧ 10 мкМ омега-3, омега-6 и омега-9 Tm и ΔG0 составили 326.2 и 87, 319.07 и 87 и 313.23 К и 86 кДж/моль соответственно. Такое же снижение свободной энергии Гиббса, которая является критерием стабильности белка, достигнуто при химической денатурации мочевиной в присутствии омега 3-6-9 жирных кислот. Взаимодействие омега 3-6-9 жирных кислот с САЧ подтверждено после сравнения с результатами ELISAанализа с L-тироксином. Хотя оценка регулярной вторичной структуры САЧ с использованием КД показала незначительные изменения после инкубации с омега 3-6-9 жирными кислотами, его третичная структура выявила наблюдаемые колебания. Таким образом, взаимодействие омега 3-6-9 жирных кислот с САЧ приводит к нестабильности и частичным структурным изменениям. Кроме того, согласно результатам молекулярного докинга, аффинность связывания омега-3, омега-6 и омега-9 с субдоменом ІІA САЧ выше, чем с субдоменом ІІIA. 

The interaction between human serum albumin (HSA) and omega 3-6-9 fatty acids (omega-3, omega-6, and omega-9), as unsaturated fatty acids, has been investigated using various methods including UV-Vis spectrophotometry, circular dichroism (CD) spectroscopy, ELISA, lifetime and fluorescence anisotropy measurements, and the visual molecular dynamics (MD) simulation. The thermodynamic parameters of HSA thermal and chemical denaturation were assessed with and without omega 3-6-9 fatty acids. The Tm and ΔG0 (298K) of sole HSA were 327.7 K and 88 kJ/mol respectively. These figures for HSA treatment with 10 µM omega-3, omega-6, and omega-9 were 326.2 K and 87 kJ/mol, 319.07 K and 87 kJ/mol, and 313.23 K and 86 kJ/mol respectively. The same manner of reduction in Gibbs free energy, which is a protein stability criterion, was achieved in chemical denaturation by urea in presence of omega 3-6-9 fatty acids. The interaction of omega 3-6-9 fatty acids with HSA was confirmed after comparing it with L-thyroxin through ELISA assay. Althoug, evaluation of the regular secondary structure of HSA using CD showed a minor change after incubation with omega 3-6-9 fatty acids, its tertiary structure revealed an observable fluctuation. Thus, it seems that the interaction of omega 3-6-9 fatty acids with HSA leads to instability and partial structural changes. Furthermore, the molecular docking results indicated that the binding affinity of omega-3, omega6, and omega-9 to subdomain ІІA of HSA was higher than subdomain ІІIA. These results provide valuable insights into the binding mechanism of omega 3-6-9 fatty acids to HSA, which could play an important role in medicinal drug delivery. 

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The authors declare that there are no conflicts of interest present.